WO2005121138A2 - Heterotricyclic compounds for use as hcv inhibitors - Google Patents

Heterotricyclic compounds for use as hcv inhibitors Download PDF

Info

Publication number
WO2005121138A2
WO2005121138A2 PCT/US2005/019499 US2005019499W WO2005121138A2 WO 2005121138 A2 WO2005121138 A2 WO 2005121138A2 US 2005019499 W US2005019499 W US 2005019499W WO 2005121138 A2 WO2005121138 A2 WO 2005121138A2
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
aryl
heteroaryl
halo
alkoxy
Prior art date
Application number
PCT/US2005/019499
Other languages
French (fr)
Other versions
WO2005121138A3 (en
Inventor
Thota Sambaiah
Singh Rajinder
Guy Laidig
Lu Henry
Original Assignee
Rigel Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rigel Pharmaceuticals, Inc. filed Critical Rigel Pharmaceuticals, Inc.
Publication of WO2005121138A2 publication Critical patent/WO2005121138A2/en
Publication of WO2005121138A3 publication Critical patent/WO2005121138A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
    • C07D471/14—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/14—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D495/14—Ortho-condensed systems

Definitions

  • HCV hepatitis C virus
  • Hepatitis C causes an estimated 8,000 to 10,000 deaths annually in the United States.
  • hepatitis C A distinct and major characteristic of hepatitis C is its tendency to cause chronic liver disease. At least 75 percent of patients with acute hepatitis C ultimately develop chronic infection, and most of these patients have accompanying chronic liver disease.
  • Chronic hepatitis C varies greatly in its course and outcome. At one end of the spectrum are patients who have no signs or symptoms of liver disease and completely normal levels of serum liver enzymes. Liver biopsy usually shows some degree of chronic hepatitis, but the degree of injury is usually mild, and the overall prognosis may be good. At the other end of the spectrum are patients with severe hepatitis C who have symptoms, HCN R ⁇ A in serum, and elevated serum liver enzymes, and who ultimately develop cirrhosis and end-stage liver disease.
  • peg-interferon alfa-2a (Pegasys®: Hoffman La Roche: Nutley, NJ) and peg-interferon alfa-2b (Pegintron®: Schering-Plough Corporation, Kenilworth, NJ). These two products are roughly equivalent in efficacy and safety, but have different dosing regimens.
  • Peg-interferon alfa-2a is given subcutaneously in a dose of 180 meg per week.
  • Peg-interferon alfa-2b is given subcutaneously weekly in doses of 1.5 meg per kilogram per week (thus in the range of 75 to 150 meg per week).
  • Ribavirin is an oral antiviral agent that has activity against a broad range of viruses. By itself, ribavirin has little effect on HCV, but adding it to interferon increases the sustained response rate by two- to three-fold. For these reasons, combination therapy is now recommended for hepatitis C and interferon monotherapy is applied only when there are specific reasons not to use ribavirin.
  • Ribavirin is an oral medication, given twice a day in 200-mg capsules for a total daily dose of 800 to 1,200 mg based upon body weight and the form of peg-interferon.
  • the recommended dose of ribavirin is 800 mg per day.
  • the dose of ribavirin is 1,000 mg for patients who weigh less than 75 kilograms (165 pounds) and 1,200 mg for those who weight more than 75 kilograms. In all situations, ribavirin is given in two divided doses daily.
  • peg-interferon alfa-2a has not been approved for use in chronic hepatitis C in the United States and is available only in clinical trials. Thus, only peg-interferon alfa-2b is available for general use.
  • Combination therapy leads to rapid improvements in serum ALT levels and disappearance of detectable HCV RNA in up to 70 percent of patients.
  • long-term improvement in hepatitis C occurs only if HCN R ⁇ A disappears during therapy and stays undetectable once therapy is stopped.
  • patients who become HCN R ⁇ A negative during treatment a proportion relapse when therapy is stopped.
  • the relapse rate is lower in patients treated with combination therapy compared with monotherapy.
  • a 48-week course of combination therapy using peg-interferon and ribavirin yields a sustained response rate of approximately 55 percent.
  • a similar course of peg-interferon monotherapy yields a sustained response rate of only 35 percent.
  • a response is considered "sustained” if HCV RNA remains undetectable for six months or more after stopping therapy.
  • the optimal duration of treatment varies depending on whether interferon monotherapy or combination therapy is used, as well as by HCV genotype. For patients treated with peg- interferon monotherapy, a 48-week course is recommended, regardless of genotype. For patients treated with combination therapy, the optimal duration of treatment depends on viral genotype. Patients with genotypes 2 and 3 have a high rate of response to combination treatment (70 to 80 percent), and a 24-week course of combination therapy yields results equivalent to those of a 48- week course. In contrast, patients with genotype 1 have a lower rate of response to combination therapy (40 to 45 percent), and a 48-week course yields a significantly better sustained response rate. Again, because of the variable responses to treatment, testing for HCV genotype is clinically useful when using combination therapy.
  • the invention provides compounds and methods for treating HCV infection.
  • the invention provides new inhibitors of HCV.
  • the invention provides compounds that are useful as inhibitors of HCV.
  • the invention provides a composition comprising an inhibitor of HCV according to the invention and a pharmaceutically acceptable carrier, excipient, or diluent.
  • the invention provides a method of inhibiting HCV in a cell, comprising contacting a cell in which inhibition of HCV is desired with an inhibitor of HCV of the invention.
  • Embodiment 1 of the invention comprises compounds of formulae I or II, ⁇ and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein, Ri is hydrogen, -OH, Ci-C ⁇ -alkyl, heterocyclyl, d-C ⁇ -al yl-OR ?
  • Y N-
  • R 8 is hydrogen or hydroxy
  • R 9 is hydroxy, Ci-C 6 -alkoxy or aryl optionally substituted with hydroxy or C ⁇ -C 6 -alkyl; or when adjacent, optionally Ri and R 2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl
  • each of Rio, R ⁇ , R ⁇ 2 and R 13 is independently selected from hydrogen, -CN, -N0 2 , -OH, d-C 6 - alkyl, aryl, C ⁇ -C 6 -alkyl-aryl, -0-halo(Ci-C 6 -alkyl), halo(Ci-C 6 -alkyl), -O-(C 3 -C 6 - cycloalkyl), -S-(C 1 -C 6 -alkyl), heterocyclyl, C ⁇ -C 6 -alkoxy, -(C ⁇
  • R 7 is hydrogen, a pro-drug group, Co-C 6 -alkyl, -(d-C ⁇ -alkyrj-OH, -(Ci-C 6 -alkyl)-0-(d-C 6 - alkyl), -(C ⁇ -C 6 -alkyl)-CN, C 2 -C 6 -alkene, heterocyclyl, aryl, heteroaryl or -(C ⁇ -C 6 -alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with d-C 6 -alkyl, nitro, hydroxy, d-C 6 -alkoxy, -CO-O-(C ⁇ -C 6 -alkyl), cyano, -O-halo(d-C 6 - alkyl), halo(Ci-C 6 -alkyl) or halogen; Ai, A 2 , A 3 , and A 4 are independently
  • a 2 are carbon, then the group formed by A ls A 2 together with Rio and R ⁇ is not phenyl;
  • a 5 is carbon or nitrogen;
  • Bi, B 2 , B 3 , and B are independently a covalent bond, C, O, -S(O)o -2 - or N, wherein when one of Bi, B , B 3 , and B is a covalent bond the R group attached to it is absent and the B ring is a five membered ring, and when B is O, -S(O)o -2 - or N then the R group attached is absent; and the dashed lines of rings A, B and C represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states; provided at least one of Bi, B , B 3 and B 4 of Formula LI is a
  • the invention provides compounds according to Formulae I and LI, wherein Bi to B 4 are carbon atoms, provided that when the compounds are according to Formulae LI and Ai, A 2 , A 3 and ⁇ are N, then Ri to R ⁇ and Rio to R ⁇ 2 are not all hydrogen; when R 2 to e are hydrogen, Ri is not -CH 3 , -CO-phenyl, -CO-H or chloro; when Ri and R 3 to Re are hydrogen, R 2 is not bromo, -CH 2 -OH, cyano, iodo, -CO-OH, -CH 3 , -CH 2 OCH 3 , morpholino, -CH 2 NH-CH 2 CH 3 , vinyl, -CH 2 CH 3 , chloro, -CH 2 NH 2 or azido; when i to R 3 and R 5 to R are hydrogen, R 4 is not chloro, ethynyl or -CH 3 ; when i to R 4 and
  • Embodiment 3 comprises compounds according to formula U
  • Ri and R 2 are independently hydrogen, -OH, C ⁇ -C 6 -alkyl, C ⁇ -C 6 -alkyl-OR 7 , C ⁇ -C 6 -alkyl-OH, aryl, heteroaryl, halogen, -CO-H, -CO-OH, -CO-OR 7 , -CO-NH 2 , -CO-NHR 7 , -CO-NR 7 R 7 , -CO-NH-OH, -CO-NH-OR 7 , -CO-NR 7 -OH, -CO-N(R 7 )-OR 7 , -CO-R 7 , C ⁇ -C 6 -alkyl-NH-OR 7 , C ⁇ -C 6 -alkyl-NH-OH, C ⁇ -C 6 -alkyl-NR 7 -OH, C ⁇ -C 6 -alkyl-NR 7 -OR 7 , C ⁇ --OR 7 , C ⁇ -C 6 -alkyl-NH-OH, C ⁇
  • Y N-
  • R 8 is hydrogen or hydroxy
  • R 9 is hydroxy, C ⁇ -C 6 -alkoxy or aryl optionally substituted with hydroxy or C ⁇ -C 6 -alkyl
  • Ri and R 2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl
  • R 3 , R 4 , R 5 and ⁇ are independently hydrogen, nitro, C ⁇ -C 6 -alkyl, aryl, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -C 6 -alkyl), -O-(C 3 -C 6 -cycloaJkyl), -S-(C ⁇ -C 6 -alkyl), -SO-(d-C 6 -alkyl), -SO 2 -(C ⁇ -C 6 - alkyl), heterocyclyl, C ⁇ -C 6 -alkoxy, -(C
  • Rio, Rn, and R ⁇ 2 are independently hydrogen, -CN, -NO 2 , -OH, C ⁇ -C 6 -alkyl, aryl, Ci-C ⁇ -alkyl- aryl, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -C 6 -alkyl), -O-(C 3 -C 6 -cycloalkyl), -S-(C ⁇ -C 6 -au yl), -SO- (C ⁇ -C 6 -alkyl), -SO 2 -(C ⁇ -C 6 -alkyl), heterocyclyl, Ci-C ⁇ -alkoxy, -(C ⁇ -C 6 -alkoxy)-aryl, azido, halogen, -OCF 3 , -CF 3 , -CO-R 7 , -CO-OH, -CO-OR 7 , -CO-NH 2 , -CO-NHR 7 , -
  • Ai, A 2 , and A 3 are independently carbon, oxygen, nitrogen, sulfur, -S(O)-, -S(O 2 )-, -C(O)-, or -N(R 7 )-;
  • a 4 is carbon or nitrogen
  • Bi, B 2 , B 3 , and B 4 are independently a covalent bond, C, O, -S(O)o- 2 - or N, wherein when one of Bi, B 2 , B 3 , and B 4 is a covalent bond the R group attached to it is absent and the B ring is a five membered ring; and the dashed lines of rings B and A represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states ;provided at least one of Bi, B 2 , B3 and B is a heteroatom.
  • Embodiment 3a of the invention comprises compounds according to formula LI wherein one of Bi, B 2 , B 3 , and B 4 is a covalent bond, and another is S and the remaining two are C, wherein the dashed lines of ring B are independently single or double bonds such that the annular S has two bonds and each annular C has four bonds; Ai, A 2 , A 3 and i are independently carbon or nitrogen; R 2 to R 6 and Rio to R ⁇ 2 are hydrogen; Ri is
  • Embodiment 3b comprises compounds according to Embodiment 3a wherein Ai to A are nitrogen.
  • the compound according to Embodiment 3b comprises compounds wherein Bi is S, B 2 and B 3 are C and B 4 is a covalent bond.
  • R 9 is methoxy.
  • heterocyclyl is pyranyl, preferably, pyran-2-yl.
  • B ring include (where the bonds in bold are shared with the C ring):
  • Embodiment 4 comprises compounds according to Embodiment 3 of the formula
  • Ri and R 2 are independently hydrogen, C ⁇ -C 6 -alkyl, heterocyclic, C ⁇ -C 6 -alkyl-OH, aryl, heteroaryl, halogen, cyano, -CO-H, -CO-OH, -CO-OR 7 , -CO-NH 2 , -CO-NHR 7 , -CO- NR 7 R 7 , -CO-NH-OH, -CO-NH-OR 7 , -CO-N(R 7 )-OR 7 , -CO-R 7 , -SO 2 NH 2 , -SO 2 NHR 7 , -SO 2 NR 7 R 7 , -SO 2 -heteroaryl, -SO 2 -aryl, -SO 3 H, -NHR 7 , C ⁇ -C 6 -alkyl-NH(R 7 )-aryl, -NH(R 7 )-aryl, -CO-heteroaryl, -NH-CO-O
  • Ri or R 2 is a group selected from
  • Ri and R 2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl;
  • R 3 , R 4 , R 5 and R 6 are independently hydrogen, nitro, C ⁇ -C 6 -alkyl, aryl, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -C 6 -alkyl), -O-(C 3 -C 6 -cycloalkyl), -S-(C C 6 -alkyl), heterocyclyl, C r C 6 - alkoxy, -(C ⁇ -C 6 -alkoxy)-aryl, azido, halogen, -OCF 3 , -CF 3 , -CO-H, -CO-OH, -CO- OR 7 , -CO-NH 2 , -CO-NHR7, -CO-NR 7 R 7 , -CO-N(R 7 )OR 7 , -CO-R 7 , -SO 3 H, -SO 2 -(d- Ce-alkyl), -SO 2 NH 2 ,
  • R 7 is hydrogen, C 0 -C 6 -alkyl, -(C ⁇ -C 6 -alkyl)-OH, -(C ⁇ -C 6 -alkyl)-O-(C ⁇ -C 6 -alkyl), -(C ⁇ -C 6 -alkyl)- CN, C 2 -C 6 -alkene, heterocyclyl, aryl, heteroaryl or -(C ⁇ -C 6 -alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with C ⁇ -C 6 - alkyl, nitro, hydroxy, C C 6 -alkoxy, -CO-O-(C ⁇ -C 6 -alkyl), cyano, -O-halo(C ⁇ -C 6 - alkyl), halo(C ⁇ -C 6 -alkyl) or halogen;
  • Ai, A 2 and A 3 are independently carbon, oxygen, or nitrogen; and ring B contains 1 or 2 nitrogen atoms;
  • Embodiment 5 comprises compounds according to Embodiment 4 of the formula
  • Ri and R 2 are independently hydrogen, Ci-C ⁇ -alkyl, heterocyclic, C ⁇ -C 6 -alkyl-OH, aryl, heteroaryl, halogen, cyano, -CO-H, -CO-OH, -CO-OR 7 , -CO-NH 2 , -CO-NHR 7 , -CO-NR 7 R 7 , -CO-NH-OH, -CO-NH-OR7, -CO-N(R 7 )-OR 7 , -CO-R 7 , -SO 3 H, -SO 2 NH 2 , -SO 2 NHR 7 , -SO 2 NR 7 R 7 , -SO 2 -heteroaryl, -SO 2 -aryl, -NHR 7 , C ⁇ -C 6 -alkyl-NH(R 7 )-aryl, -NH(R 7 )-aryl, -CO- heteroaryl, -NH-CO-O-R 7 -ary
  • R 8 is hydrogen or hydroxy;
  • Y N-, and
  • R 9 is hydroxy, Ci-d-alkoxy or aryl optionally substituted with hydroxy or Ci-C ⁇ -alkyl; or Ri and R 2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl;
  • R 3 , R 4 , R 5 and R are independently hydrogen, nitro, C ⁇ -C 6 -alkyl, aryl, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -C 6 -alkyl), -O-(C 3 -C 6 -cycloalkyl), -S-(C ⁇ -C 6 -alkyl), heterocyclyl, C ⁇ -C 6 - alkoxy, -(C ⁇ -C 6 -alkoxy)-aryl, azido, halogen, -OCF 3 , -CF 3 , -CO-H, -CO-OH, -CO- OR 7 , -CO-NH 2 , -CO-NHR7, -CO-NR 7 R 7 , -CO-N(R 7 )OR 7 , -CO-R 7 , -SO 2 -(C ⁇ -C 6 - alkyl), -SO 3 H, -SO 2
  • R 4 and R 5 together with the carbon atoms to which they are attached form a heteroaryl
  • R 7 is hydrogen, C 0 -C 6 -alkyl, -(C ⁇ -C 6 -alkyl)-OH, -(C ⁇ -C 6 -alkyl)-O-(C ⁇ -C 6 -aU yl), -(C ⁇ -C 6 -alkyl)- CN, C 2- C 6 -alkene, heterocyclyl, aryl, heteroaryl or -(C ⁇ -C 6 -alk l)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with C ⁇ -C 6 - alkyl, nitro, hydroxy, C ⁇ -C 6 -alkoxy, -CO-O-(C ⁇ -C 6 -alkyl), cyano, -O-halo(C ⁇ -C 6 - alkyl), halo(C ⁇ -C 6 -alkyl) or halogen; and provided that:
  • Ri to R are not all hydrogen; when R 2 to Re are hydrogen, Ri is not -CO-phenyl, -CO-H or chloro; when Ri and R 3 to Re are hydrogen, R 2 is not bromo, -CH 2 -OH, cyano, iodo, -CO-OH, -CH 3 , -CH 2 OCH 3 , morpholino, -CH 2 NH-CH 2 CH3, vinyl, -CH CH 3 , chloro, -CH 2 NH 2 or azido; when Ri to R 3 and R 5 to Re are hydrogen, R 4 is not chloro, ethynyl or -CH 3 ; when Ri to 4 and Re are hydrogen, R 5 is not -CH 3 or -CH 2 NH-isopropyl; when Ri to R 5 are hydrogen, Re is not -CH 2 Br, chloro, -CH 3 , -CH 2 -OH, -CO-H, -NO 2 , -NH 2 , aceta
  • Embodiment 5a of the invention comprises compounds of formula IN wherein Ri is -CO-
  • R5 is halogen
  • R 2 to R4 and Re are hydrogen. More preferrably, R 7 is hydrogen and R 5 is chloro.
  • Embodiment 5b of the invention comprises compounds of formula IN wherein Ri is aryl optionally substituted with one or more halo(C ⁇ -C 6 -alkyl), cyano, -O-halo(C ⁇ -C 6 -alkyl), or halogen, R5 is halogen, R 2 to R 4 and Re are hydrogen. More preferably, the compounds are compounds wherein Ri is phenyl optionally substituted with one or more -CF 3 or F and R 5 is chloro.
  • Embodiment 5c of the invention comprises compounds of formula IN wherein Ri is
  • R 7 is halogen and R 2 to R 5 are hydrogen.
  • Preferred compounds of this embodiment are compound wherein R7 is isobutyl, propyl, isopropyl or ethyl and R 6 is chloro.
  • Embodiment 5d of the invention comprises compounds of formula TV wherein Ri is
  • R5 is halogen
  • R 2 to R 4 and R 6 are hydrogen, preferably R 7 is tert-butyl and R 5 is chloro.
  • Embodiment 5e of the invention comprises compounds of formula IN wherein Ri is -CO-
  • Re is halo(C ⁇ -C 6 -alkyl) and R 2 to R5 are hydrogen, preferably the compound are compounds wherein R 7 is hydrogen and e is -CF 3 .
  • Embodiment 5f of the invention comprises compounds of formula IN wherein Ri is - ⁇ H-
  • R 8 is hydrogen or hydroxy
  • Y N-
  • R 9 is hydroxy or C ⁇ -C 6 -alkoxy
  • R and R 5 are independently selected from hydrogen, -S-(C ⁇ -Ce-alkyl), d-C 6 -alkyl, -O-(C 3 -C 6 - cycloalkyl), C ⁇ -C 6 -alkoxy, halogen, -SO 2 -(C ⁇ -C 6 -alkyl), heterocyclyl, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -Ce-alkyl), -NH 2 or together with the carbon atom to which they are attached form a heteroaryl group; R , R3 and Re are hydrogen; and wherein the each of
  • Embodiment 5g of the invention comprises compounds according to formula IN wherein Ri and R 2 are independently - ⁇ H(R 7 )-aryl, -CO-OH, -CO-OR 7 , C ⁇ -C 6 -alkyl, aryl or together with the carbon atoms to which they are attached form a bicyclic heteroaryl; R 3 to Re are hydrogen; and wherein each of the aryl and heteroaryl groups are optionally substituted with - CO-O-(C ⁇ -Ce-alkyl), Ci-Ce-alkyl, nitro, hydroxy, C ⁇ -C 6 -alkoxy, cyano, -O-halo(C ⁇ -C 6 -alkyl), halo(C ⁇ -Ce-alkyl) or halogen.
  • Embodiment 5h of the invention comprises compounds of formula IN wherein Ri is -CO- OH, -CO-OR 7 , or -CO- ⁇ H-OR 7 ; Re is Ci-Ce-alkyl, halogen, or -O-(C ⁇ -Ce-alkyl); and R 2 to R 5 are hydrogen.
  • Embodiment 5i of the invention comprises compounds accordmg to formula IN wherein Ri is -CO-OR 7 ; R 3 is azido or C ⁇ -C 6 -alkyl; and R 2 , to R are hydrogen.
  • Embodiment 6 comprises compounds according to formula I
  • the invention comprises a composition comprising a compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 3a-3b and 5a-5i and a pharmaceutically acceptable carrier, excipient, or diluent.
  • the invention provides a method of inhibiting HCN in a cell, comprising contacting a cell in which inhibition of HCN is desired with an inhibitor of HCV according to any one of Embodiments 1-6, 3a-3b and 5a-5i or a composition according to the second aspect of the invention. Because compounds of the invention inhibit HCV, they are also useful research tools for in vitro study HCV infections in cells and cellular systems.
  • the invention comprises a method of treating an HCV infection in a mammal, preferably a human, comprising administering to the mammal a therapeutically effective amount of a composition accordmg to the second aspect of the invention.
  • hydrocarbyl refers to a saturated, mono- or poly-unsaturated straight, branched or cyclic hydrocarbon and specifically includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, acetylenyl, propynyl, cyclopropyl, and -C ⁇ C-CH 2 (alkyl) (including -C ⁇ C-CH 2 (CH 3 ).
  • a hydrocarbyl moiety maybe defined to include a "Co-Q n - hydrocarbyl,” “Co-C n -alkyl,” or the like, in which n is an integer, as in “aryl-Co-d-alkyl.”
  • a "Co” moiety represents a direct bond. So, for example, "aryl-Co-C 3 -alkyl” encompasses both aryl-C ⁇ -C 6 -alkyl moieties as well as aryl moieties (Co-alkyl).
  • An "aryl” group is a C5-C 16 aromatic moiety comprising one to three aromatic rings, which is optionally substituted.
  • the aryl group is a C 6 -C ⁇ o aryl group.
  • Preferred aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl.
  • An "aralkyl” or “arylalkyl” group comprises an aryl group covalently linked to an alkyl group, either of which may independently be optionally substituted or unsubstituted.
  • the aralkyl group is (Ce-C ⁇ o)aryl-(C ⁇ -C 6 )alkyl, including, without limitation, benzyl, phenethyl, and naphthylmethyl.
  • heteroatom means O, S, -S(O)o-2- 5 or N.
  • Compounds that contain heteroatoms, such as -S(O)o- 2 - also comprise the chiral forms.
  • the compounds according Formulae I, LI, Ltl, and LV that contain -S(O)o- 2 - are meant to also comprise the chiral isomers.
  • the compounds according Formulae I, LI, HI, and LV comprise all types of stereochemistry, for example, racemates, enantiomers and diastereomers of the compounds.
  • the invention also includes the various regioisomers and hydro-isomers. For example, when a bond is denoted as
  • each chiral center may assume any configuration independently of the others.
  • a "heterocyclyl” group is a mono-, bi-, or tri-cyclic structure having from 3 to 16 atoms, wherein one or more annular atoms are selected from the group consisting of N, O, and S.
  • the heterocyclic group is optionally substituted on carbon at one or more positions.
  • the heterocyclic group is also independently optionally substituted on nitrogen with alkyl, aryl, aralkyl, alkylcarbonyl, alkylsulfonyl, arylcarbonyl, arylsulfonyl, alkoxycarbonyl, aralkoxycarbonyl, or on sulfur with oxo or lower alkyl.
  • Preferred heterocyclic groups include, without limitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl, and mo holino.
  • the heterocyclic group is fused to an aryl, heteroaryl, or cycloalkyl group. Examples of such fused heterocycles include, without limitation, tetrahydroquinoline and dihydrobenzofuran. Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms.
  • a particularly preferred heterocyclyl is a heteroaryl.
  • heteroaryl refers to groups having 5 to 16 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 7 ⁇ electrons shared in a cyclic array; and having, in addition to carbon atoms, from zero to three heteroatoms per ring selected from the group consisting of N, O, and S, provided there is at least one heteroatom.
  • a “heteroaralkyl” or “heteroarylalkyl” group comprises a heteroaryl group covalently linked to an alkyl group, either of which is independently optionally substituted or unsubstituted.
  • Preferred heteroalkyl groups comprise a C ⁇ -C 6 alkyl group and a heteroaryl group having 5, 6, 9, 10, 13 or 14 ring atoms. Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms.
  • Preferred heterocyclyls and heteroaryls include, but are not limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothienyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benz-lH-tetrazolyl, benz-2H-tetrazolyl, benz-3H-tetrazolyl, benz- 4H-tetrazolyl, benz-5H-tetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-l,5,2-dithiazinyl, di
  • prodrug group is a group that forms a moiety that is converted in vivo into the active compound of Formulae I and II.
  • groups are generally known in the art and include, but are not limited to, esters, alkyls, amides (-C(O)-NH-), ethers, and carbamates (-C(O 2 )-NH-).
  • prodrug groups include such groups that form, for example, amide, ether, or ester bonds or any other bond that can be hydrolyzed in vivo, for example, -C(O)-NH-O-.
  • Prodrug groups comprises any leaving groups or hydrolyzable groups that when cleaved in vivo produce the corresponding active compounds of Formulae I and JJ.
  • Open valences on the radical moieties described herein can occur on any one (or more for divalent radicals) of the atoms within the moiety.
  • the C 3 alkyl moiety includes both propyl and isopropyl.
  • a divalent C 4 alkylene moiety includes both tetramethylene (-CH 2 (CH 2 ) 2 CH 2 -) and ethylethylene (-CH(CH 2 CH 3 )CH 2 -).
  • a moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent.
  • substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluor- 3-propylphenyl.
  • substituted n-octyls include 2,4 dimethyl-5- ethyl-octyl and 3-cyclopentyl-octyl.
  • an oxo-substituted moiety is one in which both hydrogens of a methylene (-CH2-) are replaced with an oxygen to form a carbonyl (-CO-).
  • Substituents can be protected or unprotected as necessary, as known to those skilled in the art or as taught, for example, in Greene, et al, "Protective Groups in Organic Synthesis,” John Wiley and Sons, Third Edition, 1999.
  • the term pharmaceutically acceptable salt(s) refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects.
  • examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid, fn another preferred embodiment, the invention comprises the compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula -
  • Suitable substituents include, without limitation, halo, hydroxy, oxo ⁇ e.g., an annular -CH- substituted with oxo is -C(O)-) nitro, halohydrocarbyl, hydrocarbyl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, acyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido groups.
  • Preferred substituents which are themselves not further substituted (unless expressly stated otherwise) are: (a) halo, cyano, oxo, carboxy, formyl, nitro, amino, amidino, guanidino, (b) C 1 -C 5 alkyl or alkenyl or arylalkyl imino, carbamoyl, azido, carboxamido, mercapto, hydroxy, hydroxyalkyl, alkylaryl, arylalkyl, C ⁇ -C 8 alkyl, C ⁇ -C 8 alkenyl, C ⁇ -C 8 alkoxy, Ci-Cg alkoxycarbonyl, aryloxycarbonyl, C 2 -C 8 acyl, C 2 -C 8 acylamino, C ⁇ -C 8 alkylthio, arylalkylthio, arylthio, Ci-Cg alkylsulfinyl, arylalkylsulfinyl, aryl
  • halogen refers to chlorine, bromine, fluorine, or iodine.
  • halo refers to chlorine, bromine, fluorine, or iodine.
  • halo refers to chlorine, bromine, fluorine, or iodine.
  • the moiety comprises mono- to per-halogenated groups.
  • -O-halo(C ⁇ -Ce-alkyl) comprises such groups as monohaloalkoxy, dihaloalkoxy, trihaloalkoxy, etc.
  • acyl refers to an alkylcarbonyl or arylcarbonyl substituent.
  • acylamino'' refers to an amide group attached at the nitrogen atom ⁇ i.e., R-CO-NH-).
  • carbamoyl refers to an amide group attached at the carbonyl carbon atom ⁇ i.e., NH 2 -CO-).
  • the nitrogen atom of an acylamino or carbamoyl substituent is additionally substituted.
  • sulfonamido refers to a sulfonamide substituent attached by either the sulfur or the nitrogen atom.
  • amino is meant to include NH 2 , alkylamino, arylamino, and cyclic amino groups.
  • ureido as employed herein refers to a substituted or unsubstituted urea moiety.
  • radical means a chemical moiety comprising one or more unpaired electrons.
  • a moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent.
  • substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2- fluoro-3-propylphenyl.
  • substituted n-octyls include 2,4 dimethyl-5-ethyl-octyl and 3-cyclopentyl-octyl. Included within this definition are methylenes (- CH 2 -) substituted with oxygen to form carbonyl (-CO-).
  • an "unsubstituted" moiety as defined above ⁇ e.g., unsubstituted cycloalkyl, unsubstituted heteroaryl, etc.
  • moiety as defined above that does not have any of the optional substituents for which the definition of the moiety (above) otherwise provides.
  • an "aryl” includes phenyl and phenyl substituted with a halo
  • "unsubstituted aryl” does not include phenyl substituted with a halo.
  • Preferred embodiments of a particular genus of compounds of the invention include combinations of preferred embodiments.
  • the term pharmaceutically acceptable salt(s) refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects.
  • examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid.
  • inorganic acids for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like
  • organic acids such as acetic acid, oxalic acid, tart
  • the compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula NR 7 + Z " , wherein R 7 is hydrogen, alkyl, or benzyl, and Z is a counter-ion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).
  • R 7 is hydrogen, alkyl, or benzyl
  • Z is a counter-ion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate,
  • the compounds of Tables 1 to 5 are named based on the numbering system provided by ChemDraw Ultra Version 9.0.1 and by ACD Labs Version 6.0. Specifically, the numbering systems used for naming the compounds of Tables 1 to 5 are based on Structures 1 and 2 shown below.
  • the active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated.
  • the term "therapeutically effective amount” is meant to denote a dosage sufficient to inhibit proliferation of the virus in the patient.
  • a preferred dose of the active compound for all of the above-mentioned conditions is in the range from about 0.01 to 300 mg/kg, preferably 0.1 to 100 mg/kg per day, more generally 0.5 to about 25 mg per kilogram body weight of the recipient per day.
  • a typical topical dosage will range from 0.01-3% wt/wt in a suitable carrier.
  • the effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered.
  • the effective dosage can be estimated as above using the weight of the derivative, or by other means known to those skilled in the art.
  • the compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing 1 to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form.
  • An oral dosage of 1-500, preferably 10-250, more preferably 25-250 mg is usually suitable.
  • the active ingredient should be administered to achieve peak plasma concentrations of the active compound of about 0.001-30 ⁇ M, preferably about 0.01-10 ⁇ M. This maybe achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or a ⁇ iministered as a bolus of the active . ingredient.
  • the concentration of active compound in the drug composition will depend on absorption, distribution, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition.
  • the active ingredient may be administered at once, or maybe divided into a number of smaller doses to be administered at varying intervals of time.
  • Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterores; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch
  • a lubricant such as magnesium stearate or Sterores
  • a glidant such as colloidal silicon dioxide
  • dosage unit form When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil.
  • dosage unit forms can contain various other materials that modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents. See generally “Remington's Pharmaceutical Sciences,” Mack Pubhsbing Co., Easton, PA.
  • the active compound or pharmaceutically acceptable salt thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. Syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.
  • the active compound and pharmaceutically acceptable salts, hydrates, solvates and N- oxides thereof can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as antibiotics, antifungals, other anti- inflammatories, or antiviral compounds.
  • Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
  • preferred carriers are physiological saline or phosphate buffered saline (PBS).
  • the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems.
  • a controlled release formulation including implants and microencapsulated delivery systems.
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylacetic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.
  • the materials can also be obtained commercially from Alza Corporation (CA) and Gilford Pharmaceuticals (Baltimore, Md.).
  • Liposomal suspensions may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No.
  • liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidylcholine, arachadoyl phosphatidylcholine, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. Aqueous solutions of the active compound or its monophosphate, diphosphate, and/or triphosphate derivatives are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.
  • appropriate lipid(s) such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidylcholine, arachadoyl phosphatidylcholine, and cholesterol
  • compound 5 can be reacted with any suitable amine instead of H 2 N-OTHP.
  • any suitable amine instead of H 2 N-OTHP.
  • analogous chemistry can be done on heteroaryl analogs of compound 1, so long as the analogous "aniline nitrogen" in the starting compound has an adjacent CH for ring annulation as outlined below. Examples of such starting materials would include, but not be limited to, the following optionally substituted heteroaryls:
  • 2,7-Dichloroquinoline-3-carboxylic Acid (4) A dry reaction flask equipped with a magnetic stirring bar and a rubber septum was charged with 2,7-dichloro-3-formylquinoline 3 (1.1 g, 5 mmol), PDC (3.76 g, 10 mmol) and dry DMF (5 mL) and stirred at room temperature for 24 h. The reaction mixture was diluted with water (500 mL), filtered, the filtrate was saturated with NaCl and the resulting aqueous solution was extracted with EtOAc (3 x 200 mL).
  • the acid chloride was then suspended in CH 2 CI 2 (5 mL), cooled to 0 °C, to it was added pyridine (0.033 mL, 0.4 mmol) followed by H 2 N-OTHP (0.035 g, 0.3 mmol), and stirred at room temperature for 24h.
  • CH2CI 2 solution was washed with water, dried and solvent was evaporated.
  • the resulting residue was filtered through a pad of silica gel to obtain the requisite oxamic acid ester 6.
  • Example 3 The compounds of the invention according to formula V can be synthesized according to the scheme III presented below using methods well known to those skilled in the art.
  • One of ordinary skill in the art would recognize that analogous chemistry can be done on other fused heteroaryls analogous to compound la (below), so long as the nitrile- functionality can be installed on the "A" ring as outlined in Scheme III.
  • 2-halo (or other leaving group- containing) pyridines are commercially available and analogous heterocycles can be synthesized as outlined in Scheme II above using as starting materials, for example those listed above that are analogous to compound 8.
  • compounds 3a, 4a, 5a 6a and 7a are depicted as the corresponding 3H-tetrazole.
  • Assay Example 1 HCVReplicon Assay
  • Actively dividing 5-2Luc replicon cells were seeded at the density of 5000-7500 cells/well in the volume of 90 ⁇ l/well into 96 well plate(s). The cells were then incubated at 37°C and 5% CO 2 for 24 hours.
  • the 5-2 cells are replicon cells licensed from Ralf Bartenschlager (Germany) and have a self-replicating RNA molecule in the Huh7 cell; the RNA contains HCV non-structural proteins that make the self-replication possible.
  • CMV-Luc cells Actively dividing CMV-Luc cells (Luc cells in which DNA construct (CMV promoter followed by Luciferase gene) is permanently integrated into the chromosome of Huh7 cells) were seeded at the density of 5000-7500 cells/well in the volume of 90 ⁇ l/well into 96 well plate(s). The cells were then incubated at 37°C and 5% CO 2 for 24 hours. [0088] Various concentrations of compounds (in the volume of 10 ⁇ l) were added into each well 24 hours after seeding the cells. The cells were incubated with the compounds for another 24 hours before luciferase assay.
  • Protein samples were prepared from the cultured cells and resolved on a SDS-PAGE gel. [0094] After electrophoresis, the protein samples on the SDS-PAGE gel were transferred to a nitrocellulose membrane.
  • TaqMan® (Roche Molecular Systems) one step RT-PCR was performed using the RNA samples according to the manufacturer's manual. Briefly, properly diluted RNA sample, upstream primer, downstream primer, FAM-labeled probe oligo were mixed and water was added to make up the volume to 25 ⁇ l. Equal volume of 2X TaqMan Master Mix were added and the reaction was performed in an ABI Prism 7700 Sequence Detector (Applied Biosystems).

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Communicable Diseases (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention comprises tetrazoloquinine-compounds that are inhibitors of HCV. Compositions comprising the compounds in combination with a pharmaceutically acceptable carrier are also disclosed, as are methods of using the compounds and compositions to inhibit HCV infection of a cell, particular in the form of treating HCV infection in a mammal.

Description

HETEROTRICYCLIC COMPOUNDS FOR USE AS HCN INHIBITORS
Cross Reference to Related Applications [0001] This application claims priority from U.S. Provisional Patent Application No. 60/645,212, filed on January 18, 2005, from U.S. Provisional Patent AppHcation No. 60/576,535, filed on June 3, 2004, and from U.S. Provisional Patent Application No. 60/576,521, filed on June 3, 2004. Field of the Invention [0002] The present invention is in the field of small molecule inhibitors of HCV and methods of using them to inhibit HCV. Summary of the Related Art [0003] The hepatitis C virus (HCV) is one of the most important causes of chronic liver disease in the United States. It accounts for about 15 percent of acute viral hepatitis, 60 to 70 percent of chronic hepatitis, and up to 50 percent of cirrhosis, end-stage liver disease, and liver cancer. Almost 4 million Americans, or 1.8 percent of the U.S. population, have antibody to HCV (anti- HCN), indicating ongoing or previous infection with the virus. Hepatitis C causes an estimated 8,000 to 10,000 deaths annually in the United States.
[0004] A distinct and major characteristic of hepatitis C is its tendency to cause chronic liver disease. At least 75 percent of patients with acute hepatitis C ultimately develop chronic infection, and most of these patients have accompanying chronic liver disease. [0005] Chronic hepatitis C varies greatly in its course and outcome. At one end of the spectrum are patients who have no signs or symptoms of liver disease and completely normal levels of serum liver enzymes. Liver biopsy usually shows some degree of chronic hepatitis, but the degree of injury is usually mild, and the overall prognosis may be good. At the other end of the spectrum are patients with severe hepatitis C who have symptoms, HCN RΝA in serum, and elevated serum liver enzymes, and who ultimately develop cirrhosis and end-stage liver disease. In the middle of the spectrum are many patients who have few or no symptoms, mild to moderate elevations in liver enzymes, and an uncertain prognosis. Researchers estimate that at least 20 percent of patients with chronic hepatitis C develop cirrhosis, a process that takes 10 to 20 years. After 20 to 40 years, a smaller percentage of patients with chronic disease develop liver cancer. The therapy of chronic hepatitis C has evolved steadily since alpha interferon was first approved for use in this disease more than ten years ago. At the present time, the optimal regimen appears to be a 24- or 48-week course of the combination of pegylated alpha interferon and ribavirin. [0006] Two forms of peg-interferon have been developed and studied in large clinical trials: peg- interferon alfa-2a (Pegasys®: Hoffman La Roche: Nutley, NJ) and peg-interferon alfa-2b (Pegintron®: Schering-Plough Corporation, Kenilworth, NJ). These two products are roughly equivalent in efficacy and safety, but have different dosing regimens. Peg-interferon alfa-2a is given subcutaneously in a dose of 180 meg per week. Peg-interferon alfa-2b is given subcutaneously weekly in doses of 1.5 meg per kilogram per week (thus in the range of 75 to 150 meg per week).
[0007] Ribavirin is an oral antiviral agent that has activity against a broad range of viruses. By itself, ribavirin has little effect on HCV, but adding it to interferon increases the sustained response rate by two- to three-fold. For these reasons, combination therapy is now recommended for hepatitis C and interferon monotherapy is applied only when there are specific reasons not to use ribavirin.
[0008] Ribavirin is an oral medication, given twice a day in 200-mg capsules for a total daily dose of 800 to 1,200 mg based upon body weight and the form of peg-interferon. When combined with peg-interferon alfa-2b, the recommended dose of ribavirin is 800 mg per day. When combined with peg-interferon alfa-2a, the dose of ribavirin is 1,000 mg for patients who weigh less than 75 kilograms (165 pounds) and 1,200 mg for those who weight more than 75 kilograms. In all situations, ribavirin is given in two divided doses daily. [0009] At the present, peg-interferon alfa-2a has not been approved for use in chronic hepatitis C in the United States and is available only in clinical trials. Thus, only peg-interferon alfa-2b is available for general use.
[0010] Combination therapy leads to rapid improvements in serum ALT levels and disappearance of detectable HCV RNA in up to 70 percent of patients. However, long-term improvement in hepatitis C occurs only if HCN RΝA disappears during therapy and stays undetectable once therapy is stopped. Among patients who become HCN RΝA negative during treatment, a proportion relapse when therapy is stopped. The relapse rate is lower in patients treated with combination therapy compared with monotherapy. Thus, a 48-week course of combination therapy using peg-interferon and ribavirin yields a sustained response rate of approximately 55 percent. A similar course of peg-interferon monotherapy yields a sustained response rate of only 35 percent. A response is considered "sustained" if HCV RNA remains undetectable for six months or more after stopping therapy.
[0011] The optimal duration of treatment varies depending on whether interferon monotherapy or combination therapy is used, as well as by HCV genotype. For patients treated with peg- interferon monotherapy, a 48-week course is recommended, regardless of genotype. For patients treated with combination therapy, the optimal duration of treatment depends on viral genotype. Patients with genotypes 2 and 3 have a high rate of response to combination treatment (70 to 80 percent), and a 24-week course of combination therapy yields results equivalent to those of a 48- week course. In contrast, patients with genotype 1 have a lower rate of response to combination therapy (40 to 45 percent), and a 48-week course yields a significantly better sustained response rate. Again, because of the variable responses to treatment, testing for HCV genotype is clinically useful when using combination therapy.
[0012] In view of the foregoing, there is a desire for alternative, more effective agents for treating HCV infection.
SUMMARY OF THE INVENTION
[0013] The invention provides compounds and methods for treating HCV infection. The invention provides new inhibitors of HCV.
[0014] In a first aspect, the invention provides compounds that are useful as inhibitors of HCV.
[0015] In a second aspect, the invention provides a composition comprising an inhibitor of HCV according to the invention and a pharmaceutically acceptable carrier, excipient, or diluent.
[0016] In a third aspect, the invention provides a method of inhibiting HCV in a cell, comprising contacting a cell in which inhibition of HCV is desired with an inhibitor of HCV of the invention.
[0017] The foregoing merely summarizes certain aspects of the invention and is not intended to be limiting in nature. These aspects and other aspects and embodiments are described more fully below.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0018] The Embodiment 1 of the invention comprises compounds of formulae I or II,
Figure imgf000005_0001
π and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein, Ri is hydrogen, -OH, Ci-Cδ-alkyl, heterocyclyl, d-Cβ-al yl-OR?, Cι-C6-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, -CO-R7, Ci-Cβ- alkyl-NH-OR7, d-Ce-alkyl-NH-OH, C1-C6-alkyl-NR7-OH, C1-C6-alkyl-NR7-OR7, C1-C6-alkyl-CO-NHOR7, Ci-C6-a.kyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Ci-Cβ-alkyl- CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2Cl, -NHR7, C1.C6-ajJkyl-NH(R7)-aryL Cι-C6-alkyl-NR7-OR7, -NH(R7)- aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO- NH-(C1-C6-alkyl) or -NH-CO- Q-Ce-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Ci- C6-alkoxy, -CO-O-(C1-C6-akyl), cyano, -O-halofCrCβ-alkyl), halo(Cι-C6-alkyl) or halogen; each of R , R3, Rj, R5 and Re is independently selected from hydrogen, nitro, -OH, Ci-Cβ-alkyl, heterocyclyl, -O-halo(C1-C6-alkyl), halo(C1-C6-alkyl), Ci-Cβ-alkyl-ORy, d-Ce-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -OCF3, -CF , azido, -O-(C3-C6-cycloalkyl), - S-(C1-C6-alkyl), -Ce-alkoxy, -(C1-C6-alkoxy)-aryl, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, - CO-R7, d-Ce-alkyl-NH-OR?, Cι-C6-alkyl-NH-OH, C1-C6-alkyl-NR7-OH, d-Ce-alkyl-NR?- OR7, C1-C6-alkyl-CO-NHOR7, Q-Ce-alkyl-CO-NE^OR?, Cι-C6-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2- heteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2Cl, -NH2, -NHR7, -NR7R7, Q-Ce-alkyl-NHCR?)- aryl, C1-C6-alkyl-NR7-OR7, -NH(R7)-aryl, -CO-heteroaryl, -N(R7)-CO-R7, -NH-SO2-R7, - N(R7)-CO-NR7R7, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NR7-CO-OR7, -NH-CO-OR7, -NH-CO-NH-(C,-C6-alkyl) or -NH-CO-(C1-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with d-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-0-(C1-C6-akyl), cyano, -O-halo(C1-C6-alkyl), halo(d- C6-alkyl) or halogen; or any adjacent two of R3, R^ R5 and R6 together with the carbon atoms to which they are attached form an aryl, heterocyclyl or heteroaryl; or Ri and R2 are independently H or a group selected from
Figure imgf000006_0001
wherein X is =NH or =S, Y is =N-, R8 is hydrogen or hydroxy, R9 is hydroxy, Ci-C6-alkoxy or aryl optionally substituted with hydroxy or Cι-C6-alkyl; or when adjacent, optionally Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl; each of Rio, Rπ, Rι2 and R13 is independently selected from hydrogen, -CN, -N02, -OH, d-C6- alkyl, aryl, Cι-C6-alkyl-aryl, -0-halo(Ci-C6-alkyl), halo(Ci-C6-alkyl), -O-(C3-C6- cycloalkyl), -S-(C1-C6-alkyl), heterocyclyl, Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-R7, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-NR7OH, -CO-NHOR7, d-Q-alkyl-CO-NHOR?, d-Q-alkyl-CO-NRjOR?, Q-Q-alkyl-CO-NHOH, d-C6-alkyl-CO-NR7OH, -CO- N(R7)OH, -CO-NHOH, -CO-H, -SO2-R7, -SO-R7, -SO-(Cι-C6-alkyl), -SO2-(Cι-C6-alkyl), - SO2NH2, -S02NHR7, -SO2NR7R7, -CO-heteroaryl, Cι-C6-alkyl-NH-OR7, d-C6-alkyl-NH- OH C1-C6-alkyl-NR7-OH C1-C6-alkyl-NR7-OR7j -NH2, -NHR7, -NR7R7, -N(R7)-CO-R7, - NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7, or when the annular B ring atom to which it is bound is N, R3 is optionally absent, wherein each of the alkyl, alkoxy, aryl, heteroaryl and heterocyclyl are optionally substituted with one or more halogen, d-C6-alkyl, d-C6- alkoxy, -OCF3, -CF3, -CN, -NH2, -NO2, -OH, mono- or di-d-C6-alkylamino, or oxo;
R7 is hydrogen, a pro-drug group, Co-C6-alkyl, -(d-Cδ-alkyrj-OH, -(Ci-C6-alkyl)-0-(d-C6- alkyl), -(Cι-C6-alkyl)-CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with d-C6-alkyl, nitro, hydroxy, d-C6-alkoxy, -CO-O-(Cι-C6-alkyl), cyano, -O-halo(d-C6- alkyl), halo(Ci-C6-alkyl) or halogen; Ai, A2, A3, and A4 are independently carbon, oxygen, nitrogen, sulfur, -S(O)-, -S(O2)-, -C(O)-, or -N(R7)-, wherem when A! and A2 are carbon, then the group formed by Als A2 together with Rio and Rπ is not phenyl; A5 is carbon or nitrogen; Bi, B2, B3, and B are independently a covalent bond, C, O, -S(O)o-2- or N, wherein when one of Bi, B , B3, and B is a covalent bond the R group attached to it is absent and the B ring is a five membered ring, and when B is O, -S(O)o-2- or N then the R group attached is absent; and the dashed lines of rings A, B and C represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states; provided at least one of Bi, B , B3 and B4 of Formula LI is a heteroatom. [0019] In Embodiment 2, the invention provides compounds according to Formulae I and LI, wherein Bi to B4 are carbon atoms, provided that when the compounds are according to Formulae LI and Ai, A2, A3 and ι are N, then Ri to Rδ and Rio to Rι2 are not all hydrogen; when R2 to e are hydrogen, Ri is not -CH3, -CO-phenyl, -CO-H or chloro; when Ri and R3 to Re are hydrogen, R2 is not bromo, -CH2-OH, cyano, iodo, -CO-OH, -CH3, -CH2OCH3, morpholino, -CH2NH-CH2CH3, vinyl, -CH2CH3, chloro, -CH2NH2 or azido; when i to R3 and R5 to R are hydrogen, R4 is not chloro, ethynyl or -CH3; when i to R4 and R6 are hydrogen, R5 is not -CH3 or -CH2NH-isopropyl; when Ri to R5 are hydrogen, Re is not -CH2Br, chloro, -CH3, -CH2-OH, -CO-H, -NO2, -NH2, acetamido, -CH2OCH3, -CH2-CN, propyl, iodo or -CH2OCH2CH3; and the compounds of formula II are not one of the following combinations when Bι-B4 are all C:
Figure imgf000007_0001
Figure imgf000008_0002
[0020] Embodiment 3 comprises compounds according to formula U
Figure imgf000008_0001
and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein Ri and R2 are independently hydrogen, -OH, Cι-C6-alkyl, Cι-C6-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, halogen, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, -CO-R7, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH-OH, Cι-C6-alkyl-NR7-OH, Cι-C6-alkyl-NR7-OR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, d-C6-alkyl-CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2Cl, -SO2NHR7, -SO2N(R7), -NHR7, Cι.C6-alkyl-NH(R7)-aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)-aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO-NH-(Cι-C6-alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Ci-C6-a]kyl), halo(Cι-C6- alkyl) or halogen; or one of Ri or R2 is H and the other is
Figure imgf000009_0001
wherein X is =NH or =S, Y is =N-, R8 is hydrogen or hydroxy, R9 is hydroxy, Cι-C6-alkoxy or aryl optionally substituted with hydroxy or Cι-C6-alkyl; or Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl; R3, R4, R5 and δ are independently hydrogen, nitro, Cι-C6-alkyl, aryl, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl), -O-(C3-C6-cycloaJkyl), -S-(Cι-C6-alkyl), -SO-(d-C6-alkyl), -SO2-(Cι-C6- alkyl), heterocyclyl, Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO- H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-R7, -CO- NHOH, -CO-NHOR7, -CO-NR7OH, -CO-NR7OR7, -SO2-(Cι-C6-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, -NH2, -NHR7, -NR7R7, -OH, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7) -N(R7)-CO-NR7R7, or, when the annular B ring atom to which it is bound is N, R3-Rδ is optionally absent; or R and R5 together with the carbon atoms to which they are attached form a heteroaryl; R7 is hydrogen, a pro-dr g group, C0-C6-alkyl, -(Cι-C6-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6- alkyl), -(Cι-C6-alkyl)-CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or-(Cι-C6-alkyl)- aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-C6-alkyl), cyano, -O-halo(Cι- C6-alkyl), halo(Cι-C6-alkyl) or halogen;
Rio, Rn, and Rι2 are independently hydrogen, -CN, -NO2, -OH, Cι-C6-alkyl, aryl, Ci-Cδ-alkyl- aryl, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl), -O-(C3-C6-cycloalkyl), -S-(Cι-C6-au yl), -SO- (Cι-C6-alkyl), -SO2-(Cι-C6-alkyl), heterocyclyl, Ci-Cδ-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-R7, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-NHOR7; Cι-C6-aUcyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -CO-N(R7)OH, -CO-NHOH, -CO-H, -SO2-R7, -SO-R7, -SO2-(Cι-C6-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH-OH Cι-C6-alkyl-NR7-OH; Cι-C6-alkyl-NR7-OR7; -NH2, -NHR7, -NR7R7, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7, wherein each of the alkyl, alkoxy, aryl, heteroaryl and heterocyclyl are optionally substituted with one or more halogen, Cι-C6-alkyl, Cι-C6-alkoxy, -OCF3, -CF3, -CN, -NH2, -NO2, -OH, mono- or di-Cι-C6-alkylamino, or oxo;
Ai, A2, and A3 are independently carbon, oxygen, nitrogen, sulfur, -S(O)-, -S(O2)-, -C(O)-, or -N(R7)-;
A4 is carbon or nitrogen;
Bi, B2, B3, and B4 are independently a covalent bond, C, O, -S(O)o-2- or N, wherein when one of Bi, B2, B3, and B4 is a covalent bond the R group attached to it is absent and the B ring is a five membered ring; and the dashed lines of rings B and A represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states ;provided at least one of Bi, B2, B3 and B is a heteroatom.
[0021] Embodiment 3a of the invention comprises compounds according to formula LI wherein one of Bi, B2, B3, and B4 is a covalent bond, and another is S and the remaining two are C, wherein the dashed lines of ring B are independently single or double bonds such that the annular S has two bonds and each annular C has four bonds; Ai, A2, A3 and i are independently carbon or nitrogen; R2 to R6 and Rio to Rι2 are hydrogen; Ri is
Figure imgf000011_0001
wherein R9 is hydroxy or Cι-C3-alkoxy, -CO-H, -CO-OH, -CO-NH-OH, or -CO-NH-OR7, wherein R is a heterocyclyl, wherein the heterocyclyl groups is optionally substituted with Ci- Cβ-alkyl, hydroxy, Cι-C6-alkoxy, cyano, or halogen. [0022] Embodiment 3b comprises compounds according to Embodiment 3a wherein Ai to A are nitrogen. Preferably, the compound according to Embodiment 3b comprises compounds wherein Bi is S, B2 and B3 are C and B4 is a covalent bond. Preferably, R9 is methoxy. Also preferred are compounds according to Embodiment 3a wherein the heterocyclyl is pyranyl, preferably, pyran-2-yl. [0023] Illustrative, but non-limiting, examples of the B ring include (where the bonds in bold are shared with the C ring):
Figure imgf000011_0002
[0024] The following table presents preferred embodiments of Bι-B4:
Figure imgf000011_0005
Figure imgf000011_0003
Figure imgf000011_0004
[0025] Embodiment 4 comprises compounds according to Embodiment 3 of the formula
Figure imgf000012_0001
and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein
Ri and R2 are independently hydrogen, Cι-C6-alkyl, heterocyclic, Cι-C6-alkyl-OH, aryl, heteroaryl, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO- NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-N(R7)-OR7, -CO-R7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -SO3H, -NHR7, Cι-C6-alkyl-NH(R7)-aryl, -NH(R7)-aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SOa-aryl, -NH-CO- OR7, -NH-CO-NH-(Cι-C6-alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Ci-d-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl) or halogen; or
Ri or R2 is a group selected from
Figure imgf000012_0002
wherein R8 is hydrogen or hydroxy; X is =NH or =S, Y is =N-, and R9 is hydroxy, Cι-C6-alkoxy or aryl optionally substituted with hydroxy or Cι-C6-alkyl; or
Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl;
R3, R4, R5 and R6 are independently hydrogen, nitro, Cι-C6-alkyl, aryl, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl), -O-(C3-C6-cycloalkyl), -S-(C C6-alkyl), heterocyclyl, CrC6- alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-OH, -CO- OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-R7, -SO3H, -SO2-(d- Ce-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, -NH2, -NHR7, -NR7R7, -OH, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7; R4 and R5 together with the carbon atoms to which they are attached form a heteroaryl;
R7 is hydrogen, C0-C6-alkyl, -(Cι-C6-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6-alkyl), -(Cι-C6-alkyl)- CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Cι-C6- alkyl, nitro, hydroxy, C C6-alkoxy, -CO-O-(Cι-C6-alkyl), cyano, -O-halo(Cι-C6- alkyl), halo(Cι-C6-alkyl) or halogen;
Ai, A2 and A3 are independently carbon, oxygen, or nitrogen; and ring B contains 1 or 2 nitrogen atoms;
[0026] Embodiment 5 comprises compounds according to Embodiment 4 of the formula
Figure imgf000013_0001
or a pharmaceutically acceptable salt thereof wherein
Ri and R2 are independently hydrogen, Ci-Cδ-alkyl, heterocyclic, Cι-C6-alkyl-OH, aryl, heteroaryl, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-N(R7)-OR7, -CO-R7, -SO3H, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -NHR7, Cι-C6-alkyl-NH(R7)-aryl, -NH(R7)-aryl, -CO- heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO-NH-(Cι-C6- alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl) or halogen; or Ri or R2 is a group selected from
Figure imgf000013_0002
wherein R8 is hydrogen or hydroxy; X is =NH or =S; Y is =N-, and R9 is hydroxy, Ci-d-alkoxy or aryl optionally substituted with hydroxy or Ci-Cβ-alkyl; or Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl;
R3, R4, R5 and R are independently hydrogen, nitro, Cι-C6-alkyl, aryl, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl), -O-(C3-C6-cycloalkyl), -S-(Cι-C6-alkyl), heterocyclyl, Cι-C6- alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-OH, -CO- OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-R7, -SO2-(Cι-C6- alkyl), -SO3H, -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, -NH2, -NHR7, -NR7R7, -OH, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7;
R4 and R5 together with the carbon atoms to which they are attached form a heteroaryl;
R7 is hydrogen, C0-C6-alkyl, -(Cι-C6-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6-aU yl), -(Cι-C6-alkyl)- CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alk l)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Cι-C6- alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-C6-alkyl), cyano, -O-halo(Cι-C6- alkyl), halo(Cι-C6-alkyl) or halogen; and provided that:
Ri to R are not all hydrogen; when R2 to Re are hydrogen, Ri is not -CO-phenyl, -CO-H or chloro; when Ri and R3 to Re are hydrogen, R2 is not bromo, -CH2-OH, cyano, iodo, -CO-OH, -CH3, -CH2OCH3, morpholino, -CH2NH-CH2CH3, vinyl, -CH CH3, chloro, -CH2NH2 or azido; when Ri to R3 and R5 to Re are hydrogen, R4 is not chloro, ethynyl or -CH3; when Ri to 4 and Re are hydrogen, R5 is not -CH3 or -CH2NH-isopropyl; when Ri to R5 are hydrogen, Re is not -CH2Br, chloro, -CH3, -CH2-OH, -CO-H, -NO2, -NH2, acetamido, -CH2OCH3, -CH2-CN, propyl, iodo or -CH2OCH2CH3; and the compounds of formula IN are not one of the following combinations
Figure imgf000014_0001
Figure imgf000015_0001
[0027] Embodiment 5a of the invention comprises compounds of formula IN wherein Ri is -CO-
ΝH-OR7, R5 is halogen, R2 to R4 and Re are hydrogen. More preferrably, R7 is hydrogen and R5 is chloro.
[0028] Embodiment 5b of the invention comprises compounds of formula IN wherein Ri is aryl optionally substituted with one or more halo(Cι-C6-alkyl), cyano, -O-halo(Cι-C6-alkyl), or halogen, R5 is halogen, R2 to R4 and Re are hydrogen. More preferably, the compounds are compounds wherein Ri is phenyl optionally substituted with one or more -CF3 or F and R5 is chloro.
[0029] Embodiment 5c of the invention comprises compounds of formula IN wherein Ri is
-CO-ΝH-OR7, Re is halogen and R2 to R5 are hydrogen. Preferred compounds of this embodiment are compound wherein R7 is isobutyl, propyl, isopropyl or ethyl and R6 is chloro.
[0030] Embodiment 5d of the invention comprises compounds of formula TV wherein Ri is
-NH-CO-OR7, R5 is halogen, R2 to R4 and R6 are hydrogen, preferably R7 is tert-butyl and R5 is chloro.
[0031] Embodiment 5e of the invention comprises compounds of formula IN wherein Ri is -CO-
OR7, Re is halo(Cι-C6-alkyl) and R2 to R5 are hydrogen, preferably the compound are compounds wherein R7 is hydrogen and e is -CF3.
[0032] Embodiment 5f of the invention comprises compounds of formula IN wherein Ri is -ΝH-
CO-ΝH-SO2-aryl, -NH-CO-NH-(Cι-C6-alkyl), Cι-C6-alkyl-OH, -CO-H, -CO-NH2, -CO-NH-R7,
-CO-NH-OR7, -SO2-aryl, -SO2-heteroaryl, heteroaryl, -CO-OR or a group represented by
Figure imgf000016_0001
wherein R8 is hydrogen or hydroxy, X is =N or =S, Y is =N-, and R9 is hydroxy or Cι-C6-alkoxy; R and R5 are independently selected from hydrogen, -S-(Cι-Ce-alkyl), d-C6-alkyl, -O-(C3-C6- cycloalkyl), Cι-C6-alkoxy, halogen, -SO2-(Cι-C6-alkyl), heterocyclyl, -O-halo(Cι-C6-alkyl), halo(Cι-Ce-alkyl), -NH2 or together with the carbon atom to which they are attached form a heteroaryl group; R , R3 and Re are hydrogen; and wherein the each of the aryl, heteroaryl and heterocyclyl groups are optionally substituted with -CO-O-(Cι-C6-alkyl), Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, cyano, -O-halo(Cι-Ce-alkyl), halo(Cι-Ce-alkyl) or halogen. [0033] Embodiment 5g of the invention comprises compounds according to formula IN wherein Ri and R2 are independently -ΝH(R7)-aryl, -CO-OH, -CO-OR7, Cι-C6-alkyl, aryl or together with the carbon atoms to which they are attached form a bicyclic heteroaryl; R3 to Re are hydrogen; and wherein each of the aryl and heteroaryl groups are optionally substituted with - CO-O-(Cι-Ce-alkyl), Ci-Ce-alkyl, nitro, hydroxy, Cι-C6-alkoxy, cyano, -O-halo(Cι-C6-alkyl), halo(Cι-Ce-alkyl) or halogen.
[0034] Embodiment 5h of the invention comprises compounds of formula IN wherein Ri is -CO- OH, -CO-OR7, or -CO-ΝH-OR7; Re is Ci-Ce-alkyl, halogen, or -O-(Cι-Ce-alkyl); and R2 to R5 are hydrogen.
[0035] Embodiment 5i of the invention comprises compounds accordmg to formula IN wherein Ri is -CO-OR7; R3 is azido or Cι-C6-alkyl; and R2, to R are hydrogen. [0036] Embodiment 6 comprises compounds according to formula I
Figure imgf000016_0002
wherein Ri-Re, Rιo-Ri3> A1-A5, and B1-B5 are as defined for Embodiment 1, above. [0037] fn a second aspect, the invention comprises a composition comprising a compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 3a-3b and 5a-5i and a pharmaceutically acceptable carrier, excipient, or diluent.
[0038] In a third aspect, the invention provides a method of inhibiting HCN in a cell, comprising contacting a cell in which inhibition of HCN is desired with an inhibitor of HCV according to any one of Embodiments 1-6, 3a-3b and 5a-5i or a composition according to the second aspect of the invention. Because compounds of the invention inhibit HCV, they are also useful research tools for in vitro study HCV infections in cells and cellular systems. [0039] In a preferred embodiment of the third aspect, the invention comprises a method of treating an HCV infection in a mammal, preferably a human, comprising administering to the mammal a therapeutically effective amount of a composition accordmg to the second aspect of the invention.
Definitions
[0040] Unless expressly stated to the contrary, the following definitions apply uniformly throughout. For simplicity, the substituents have been defined primarily as univalent chemical moieties {e.g., alkyl, aryl, etc.). Nevertheless, such terms are also used to convey corresponding multivalent moieties under the appropriate structural circumstances. All atoms are understood to have their normal number of valences for bond formation {i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of the S). Also, where a chemical structure exists in multiple tautomeric forms, all are envisioned as part of the invention. [0041] The term hydrocarbyl refers to a saturated, mono- or poly-unsaturated straight, branched or cyclic hydrocarbon and specifically includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, acetylenyl, propynyl, cyclopropyl, and -C≡C-CH2(alkyl) (including -C≡C-CH2(CH3). A hydrocarbyl moiety maybe defined to include a "Co-Qn- hydrocarbyl," "Co-Cn-alkyl," or the like, in which n is an integer, as in "aryl-Co-d-alkyl." In these instances a "Co" moiety represents a direct bond. So, for example, "aryl-Co-C3-alkyl" encompasses both aryl-Cι-C6-alkyl moieties as well as aryl moieties (Co-alkyl). [0042] An "aryl" group is a C5-C16 aromatic moiety comprising one to three aromatic rings, which is optionally substituted. Preferably, the aryl group is a C6-Cιo aryl group. Preferred aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl. An "aralkyl" or "arylalkyl" group comprises an aryl group covalently linked to an alkyl group, either of which may independently be optionally substituted or unsubstituted. Preferably, the aralkyl group is (Ce-Cιo)aryl-(Cι-C6)alkyl, including, without limitation, benzyl, phenethyl, and naphthylmethyl. [0043] The term heteroatom means O, S, -S(O)o-2-5 or N. Compounds that contain heteroatoms, such as -S(O)o-2-, also comprise the chiral forms. Thus, the compounds according Formulae I, LI, Ltl, and LV that contain -S(O)o-2- are meant to also comprise the chiral isomers. Further, the compounds according Formulae I, LI, HI, and LV comprise all types of stereochemistry, for example, racemates, enantiomers and diastereomers of the compounds. The invention also includes the various regioisomers and hydro-isomers. For example, when a bond is denoted as
" ", it includes " — "^ " or " "" " (that is, both S and R configurations). Further, when a compound includes one or more chiral centers, each chiral center may assume any configuration independently of the others.
[0044] A "heterocyclyl" group is a mono-, bi-, or tri-cyclic structure having from 3 to 16 atoms, wherein one or more annular atoms are selected from the group consisting of N, O, and S. The heterocyclic group is optionally substituted on carbon at one or more positions. The heterocyclic group is also independently optionally substituted on nitrogen with alkyl, aryl, aralkyl, alkylcarbonyl, alkylsulfonyl, arylcarbonyl, arylsulfonyl, alkoxycarbonyl, aralkoxycarbonyl, or on sulfur with oxo or lower alkyl. Preferred heterocyclic groups include, without limitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl, and mo holino. In certain preferred embodiments, the heterocyclic group is fused to an aryl, heteroaryl, or cycloalkyl group. Examples of such fused heterocycles include, without limitation, tetrahydroquinoline and dihydrobenzofuran. Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms. [0045] A particularly preferred heterocyclyl is a heteroaryl. As used herein, the term "heteroaryl" refers to groups having 5 to 16 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 7τ electrons shared in a cyclic array; and having, in addition to carbon atoms, from zero to three heteroatoms per ring selected from the group consisting of N, O, and S, provided there is at least one heteroatom. A "heteroaralkyl" or "heteroarylalkyl" group comprises a heteroaryl group covalently linked to an alkyl group, either of which is independently optionally substituted or unsubstituted. Preferred heteroalkyl groups comprise a Cι-C6 alkyl group and a heteroaryl group having 5, 6, 9, 10, 13 or 14 ring atoms. Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms.
[0046] Preferred heterocyclyls and heteroaryls include, but are not limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothienyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benz-lH-tetrazolyl, benz-2H-tetrazolyl, benz-3H-tetrazolyl, benz- 4H-tetrazolyl, benz-5H-tetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-l,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazoUdinyl, imidazolinyl, imidazolyl, lH-indazolyl, indolenyl, indohnyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazoh'dinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-l,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2,3-rriazolyl, 1,2,4-triazolyl, 1,2,5- triazolyl, 1,3,4-triazolyl, benzodiazepinyl, and xanthenyl.
[0047] The term "prodrug group" is a group that forms a moiety that is converted in vivo into the active compound of Formulae I and II. Such groups are generally known in the art and include, but are not limited to, esters, alkyls, amides (-C(O)-NH-), ethers, and carbamates (-C(O2)-NH-). Thus, prodrug groups include such groups that form, for example, amide, ether, or ester bonds or any other bond that can be hydrolyzed in vivo, for example, -C(O)-NH-O-. Prodrug groups comprises any leaving groups or hydrolyzable groups that when cleaved in vivo produce the corresponding active compounds of Formulae I and JJ.
[0048] Open valences on the radical moieties described herein can occur on any one (or more for divalent radicals) of the atoms within the moiety. For example, the C3 alkyl moiety includes both propyl and isopropyl. As another example, a divalent C4 alkylene moiety includes both tetramethylene (-CH2(CH2)2CH2-) and ethylethylene (-CH(CH2CH3)CH2-). [0049] A moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent. As a non-limiting example, substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluor- 3-propylphenyl. As another non-limiting example, substituted n-octyls include 2,4 dimethyl-5- ethyl-octyl and 3-cyclopentyl-octyl. As another example, an oxo-substituted moiety is one in which both hydrogens of a methylene (-CH2-) are replaced with an oxygen to form a carbonyl (-CO-).
[0050] Substituents can be protected or unprotected as necessary, as known to those skilled in the art or as taught, for example, in Greene, et al, "Protective Groups in Organic Synthesis," John Wiley and Sons, Third Edition, 1999.
[0051] As used herein, the term pharmaceutically acceptable salt(s) refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects. Examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid, fn another preferred embodiment, the invention comprises the compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula -NR + Z-, wherein R is hydrogen, alkyl, or benzyl, and Z is a counter-ion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate). For the purposes of the specification and claims, the term salt is intended to encompass complexes as well.
[0052] As employed herein, when a moiety {e.g., cycloalkyl, hydrocarbyl, aryl, heteroaryl, heterocyclic, urea, etc.) is described as "optionally substituted" it is meant that the group optionally has from one to four, preferably from one to three, more preferably one or two, non- hydrogen substituents. Suitable substituents include, without limitation, halo, hydroxy, oxo {e.g., an annular -CH- substituted with oxo is -C(O)-) nitro, halohydrocarbyl, hydrocarbyl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, acyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido groups. Preferred substituents, which are themselves not further substituted (unless expressly stated otherwise) are: (a) halo, cyano, oxo, carboxy, formyl, nitro, amino, amidino, guanidino, (b) C1-C5 alkyl or alkenyl or arylalkyl imino, carbamoyl, azido, carboxamido, mercapto, hydroxy, hydroxyalkyl, alkylaryl, arylalkyl, Cι-C8 alkyl, Cι-C8 alkenyl, Cι-C8 alkoxy, Ci-Cg alkoxycarbonyl, aryloxycarbonyl, C2-C8 acyl, C2-C8 acylamino, Cι-C8 alkylthio, arylalkylthio, arylthio, Ci-Cg alkylsulfinyl, arylalkylsulfinyl, arylsulfinyl, Cι-C8 alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, Co-Ce N-alkyl carbamoyl, d-Cis ./V'jN-dialkylcarbamoyl, C3-C7 cycloalkyl, aroyl, aryloxy, arylalkyl ether, aryl, aryl fused to a cycloalkyl or heterocycle or another aryl ring, C3-C7 heterocycle, C5-Cι heteroaryl or any of these rings fused or spiro-fused to a cycloalkyl, heterocyclyl, or aryl, wherein each of the foregoing is further optionally substituted with one more moieties listed in (a), above; and (c) -(CH2)S-NR30R31, wherein s is from 0 (in which case the nitrogen is directly bonded to the moiety that is substituted) to 6, and R30 and R31 are each independently hydrogen, cyano, oxo, carboxamido, amidino, Cι-C8 hydroxyalkyl, Cι-C3 alkylaryl, aryl-Ci-d alkyl, Cι-C8 alkyl, Cι-C8 alkenyl, Cι-C8 alkoxy, Cι-C8 alkoxycarbonyl, aryloxycarbonyl, aryl-Cι-C3 alkoxycarbonyl, d-Cs acyl, Cι-C8 alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, aroyl, aryl, cycloalkyl, heterocyclyl, or heteroaryl, wherein each of the foregoing is further optionally substituted with one more moieties listed in (a), above; or R30 and R31 taken together with the N to which they are be attached form a heterocyclyl or heteroaryl, each of which is optionally substituted with from 1 to 3 substituents from (a), above. [0053] The term "halogen" or "halo" as employed herein refers to chlorine, bromine, fluorine, or iodine. When the term "halo" is used to describe a moiety, as in, for example, -O-halo(Cι-C6- alkyl) or halo(Cι-Ce-alkyl), the moiety comprises mono- to per-halogenated groups. For example, -O-halo(Cι-Ce-alkyl) comprises such groups as monohaloalkoxy, dihaloalkoxy, trihaloalkoxy, etc. As herein employed, the term "acyl" refers to an alkylcarbonyl or arylcarbonyl substituent. The term "acylamino'' refers to an amide group attached at the nitrogen atom {i.e., R-CO-NH-). The term "carbamoyl" refers to an amide group attached at the carbonyl carbon atom {i.e., NH2-CO-). The nitrogen atom of an acylamino or carbamoyl substituent is additionally substituted. The term "sulfonamido" refers to a sulfonamide substituent attached by either the sulfur or the nitrogen atom. The term "amino" is meant to include NH2, alkylamino, arylamino, and cyclic amino groups. The term "ureido" as employed herein refers to a substituted or unsubstituted urea moiety.
[0054] The term "radical" as used herein means a chemical moiety comprising one or more unpaired electrons.
[0055] A moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent. As a non-limiting example, substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2- fluoro-3-propylphenyl. As another non-limiting example, substituted n-octyls include 2,4 dimethyl-5-ethyl-octyl and 3-cyclopentyl-octyl. Included within this definition are methylenes (- CH2-) substituted with oxygen to form carbonyl (-CO-).
[0056] An "unsubstituted" moiety as defined above {e.g., unsubstituted cycloalkyl, unsubstituted heteroaryl, etc.) means that moiety as defined above that does not have any of the optional substituents for which the definition of the moiety (above) otherwise provides. Thus, for example, while an "aryl" includes phenyl and phenyl substituted with a halo, "unsubstituted aryl" does not include phenyl substituted with a halo.
[0057] Preferred embodiments of a particular genus of compounds of the invention include combinations of preferred embodiments.
[0058] As used herein, the term pharmaceutically acceptable salt(s) refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects. Examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid. The compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula NR7 + Z", wherein R7 is hydrogen, alkyl, or benzyl, and Z is a counter-ion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).
[0059] The compounds of Tables 1 to 5 are named based on the numbering system provided by ChemDraw Ultra Version 9.0.1 and by ACD Labs Version 6.0. Specifically, the numbering systems used for naming the compounds of Tables 1 to 5 are based on Structures 1 and 2 shown below.
Figure imgf000023_0001
or Structure 1 Structure 2
[0060] The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated. The term "therapeutically effective amount" is meant to denote a dosage sufficient to inhibit proliferation of the virus in the patient. A preferred dose of the active compound for all of the above-mentioned conditions is in the range from about 0.01 to 300 mg/kg, preferably 0.1 to 100 mg/kg per day, more generally 0.5 to about 25 mg per kilogram body weight of the recipient per day. A typical topical dosage will range from 0.01-3% wt/wt in a suitable carrier. The effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered. If the derivative exhibits activity in itself, the effective dosage can be estimated as above using the weight of the derivative, or by other means known to those skilled in the art. [0061] The compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing 1 to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form. An oral dosage of 1-500, preferably 10-250, more preferably 25-250 mg is usually suitable.
[0062] The active ingredient should be administered to achieve peak plasma concentrations of the active compound of about 0.001-30 μM, preferably about 0.01-10 μM. This maybe achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or a<iministered as a bolus of the active . ingredient.
[0063] The concentration of active compound in the drug composition will depend on absorption, distribution, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or maybe divided into a number of smaller doses to be administered at varying intervals of time.
[0064] Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
[0065] The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterores; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials that modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents. See generally "Remington's Pharmaceutical Sciences," Mack Pubhsbing Co., Easton, PA. [0066] The active compound or pharmaceutically acceptable salt thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. Syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.
[0067] The active compound and pharmaceutically acceptable salts, hydrates, solvates and N- oxides thereof can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as antibiotics, antifungals, other anti- inflammatories, or antiviral compounds.
[0068] Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
[0069] If administered intravenously, preferred carriers are physiological saline or phosphate buffered saline (PBS).
[0070] In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylacetic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation (CA) and Gilford Pharmaceuticals (Baltimore, Md.). Liposomal suspensions may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidylcholine, arachadoyl phosphatidylcholine, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. Aqueous solutions of the active compound or its monophosphate, diphosphate, and/or triphosphate derivatives are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.
Synthesis [0071] The compounds of the mvention can be synthesized according to the scheme presented below using methods well known to those skilled in the art. The synthesis procedures described in the schemes below, as well as methods well known to those skilled in the art are practised using safety precautions known in the art for azide and tetrazole chemistries. One skilled in the art will recognize that the substituents of the starting compound 1 can be varied using well known synthetic procedures. For example, various types of substitution reactions can take place on the phenyl group of compound 1 to give the alkoxy, alkyl, haloalkyl, or sulfonylalkyl product. Similarly, various types of substitution reactions or condensation reactions on any of the species in Scheme 1 will result in the various compounds described in this invention. For example, compound 5 can be reacted with any suitable amine instead of H2N-OTHP. Also, one of ordinary skill in the art would recognize that analogous chemistry can be done on heteroaryl analogs of compound 1, so long as the analogous "aniline nitrogen" in the starting compound has an adjacent CH for ring annulation as outlined below. Examples of such starting materials would include, but not be limited to, the following optionally substituted heteroaryls:
Figure imgf000026_0001
Example 1
Figure imgf000027_0001
8-chloro-N-hydroxytetrazolo[l,5-a]quinoline-4-carboxamide (7) Scheme 1
Figure imgf000027_0002
[0072] 2,7-Dichloro-3-formylquinoline (3): To a dry reaction flask equipped with a reflux condenser, a magnetic stirring bar and a rubber septum with a N2 inlet was placed 3- chloroacetanilide (2) (8.45 g, 50 mmol) and POCI3 (34 mL, 365 mmol). The reaction mixture was cooled to 0 °C and to it was added DMF (10.5 mL, 135 mmol) over a period of 20 minutes. It was stirred at that temperature for 1 h and then at 65-75 °C for 24 h. The resulting slurry was cooled to room temperature, poured over a crushed-ice (1 Kg), digested for 1 h, the resulting solid was filtered, washed well with water and dried to give 5.26g (47%) of the desired 2,7- dichloro-3-formylquinoline 3. 1H NMR (CDCI3): δ 10.54 (s, 1H), 8.74 (s, 1H), 8.07 (d, 1H, J= 2.1 Hz), 7.93 (d, 1H, J= 9.0 Hz),7.61 (dd, 1H, J= 2.4 and 8.7 Hz); LCMS: 30.83 min.; MS (m/z): 226 (Nt).
[0073] 2,7-Dichloroquinoline-3-carboxylic Acid (4): A dry reaction flask equipped with a magnetic stirring bar and a rubber septum was charged with 2,7-dichloro-3-formylquinoline 3 (1.1 g, 5 mmol), PDC (3.76 g, 10 mmol) and dry DMF (5 mL) and stirred at room temperature for 24 h. The reaction mixture was diluted with water (500 mL), filtered, the filtrate was saturated with NaCl and the resulting aqueous solution was extracted with EtOAc (3 x 200 mL). The EtOAc extract was dried over anhydrous Na2SO4 and solvent was removed to give 1.02 g (85%) of the desired 2,7-dichloroquinoline-3-carboxylic acid 4. TLC Rf: 0.12 (20% MeOH/EtOAc); 1H NMR (acetone d6): 8.96 (s, 1H); 8.22 (d, 1H, J= 9.0 Hz), 8.04 (d, 1H, J= 1.8 Hz), 7.97 (bs, 1H), 7.74 (dd, 1H, J= 1.8 and 8.7 Hz); LCMS (ret. time): 25.15 min.; MS {m/z): 241 (MH4).
[0074] 8-Chlorotetrazolo[l,5-a]quinoline-4-carboxylic Acid (5): To a dry reaction vial with a screw cap was placed 2,7-dichloroquinoline-3-carboxylic acid (0.241 g, 1 mmol), sodium azide (0.078 g, 1.2 mmol) and dry DMF (3 mL) and shaken at 65-75 °C for a period of 24 h. The reaction mixture was diluted with water (100 mL), saturated with NaCl, the resulting solid was filtered and dried to give 0.100 g (41%) of the carboxytetrazole 5. TLC Rf: 0.06 (20% MeOH/EtOAc); 1H NMR (DMSO de + CDC13): δ 8.71 (bs, 1H), 8.60 (s, 1H), 8.30 (d, 1H, J= 7.8 Hz), 7.79 (d, 1H, J= 7.5 Hz); LCMS (ret. time): 21.28; MS {m/z): 249 (MH ). [0075] 8-chloro-N-(tetrahydro-2H-pyran-2-yloxy) tetrazo!o[l,5-a] quinoline-4-carboxamide (6): To a suspension of carboxytetrazole (5, 0.05 g, 0.2 mmol) in CH2C12 (5 mL) at 0 °C under N2 was added DMF (4-drops) followed by (COCl)2 (IM CH2C12, 0.15 mL, 0.3 mmol) and the reaction was stirred for lh. The reaction solvent was removed and the resulting residue of acid chloride was dried under high vacuum. The acid chloride was then suspended in CH2CI2 (5 mL), cooled to 0 °C, to it was added pyridine (0.033 mL, 0.4 mmol) followed by H2N-OTHP (0.035 g, 0.3 mmol), and stirred at room temperature for 24h. The CH2CI2 solution was washed with water, dried and solvent was evaporated. The resulting residue was filtered through a pad of silica gel to obtain the requisite oxamic acid ester 6. 1H NMR (CDC13): δ 11.58 (s, 1H), 8.92 (s, 1H), 8.74 (d, 1H, J= 1.8 Hz), 8.10 (d, 1H, J= 8.1 Hz), 7.77 (dd, 1H, J= 1.8 Hz and 8.7 Hz), 5.26 (m, 1H), 4.15 (m, 1H), 3.76 (m, 1H), 2.10-1.50 (m, 6H); LCMS (ret. time): 21.46 min.; MS (m z): 347 (MT ).
[0076] 8-chloro-N-hydroxytetrazolo[l,5-a] quinoline-4-carboxamide (7): To a solution of THP oxamate 6 (0.035 g, 0.1 mmol) in THF (1 mL) was added 2N HC1 (1 mL) and stirred at room temperature for 6h. The reaction solvent was removed under a reduced pressure and the resulting aqueous solution was diluted with water. The precipitated solid was filtered, washed with water and dried to give the desired oxamic acid 7. 1H NMR (CDC13): δ 8.80 (s, 1H), 8.64 (bd, 1H), 8.06 (d, 1H, J= 8.1 Hz), 7.70 (dd, 1H, J= 2.4 and 8.7 Hz), LCMS (ret. time): 20.47 min.; MS (m/z): 264 (MET1). Example 2
Figure imgf000029_0001
tetrazoIo[l,5-α]thieno[2,3-e]pyridine-4-carboxyIic acid (12)
Example 2a
Figure imgf000029_0002
/V-(tetrahydro-2H-pyran-2-yloxy)tetrazolo[l,5-fl]thieno[2,3-e]pyridine-4-carboxamide (13)
Example 2b
Figure imgf000029_0003
iV-hydroxytetrazolo[l,5- ]thieno[2,3-e]pyridine-4-carboxamide (14)
Scheme H
Figure imgf000030_0001
[0077] Preparation ϊE & Z of 3-acetylthiopheneoxime (9): To a solution of hydroxylamine hydrochloride {6.66 g, 0.096 mole) in water (15 mL) was added 20% NaOH solution (20 mL). The mixture was cooled in an ice-bath and 3-acetylthiophene (10 g, 0.08 mole) was added. The reaction mixture was allowed to warm to room temperature and then heated at 100°C for overnight. Upon cooling, solid separated which was collected by filtration. Recrystallized from hot water gave colorless solid 9.38 g (83%) of desired product 9.1H NMR (CDC13, 300 MHz): δ 8.65 (s, IH), 7.46 (d, IH, J= 1.5 Hz), 7.40 (d, IH, J= 1.2 Hz), 7.31 (d, IH, J= 3.0 Hz), 2.28 (s, 3H); LCMS (m/z): 142 (MH÷).
[0078] Preparation of 5-chlorothienyI[3,2-b]pridine-6-carboxaIdehyde (10): To a solution of oxime 9 (4.0 g, 28.37 mmol) in ether (40 mL), cooled in an ice/water bath was slowly added POCl3 (25.6 mL). The temperature of the reaction mixture was maintained at 0-10 °C for 2 h while DMF (4.88 mL) was slowly added. The temperature of the reaction was raised and the ether was distilled off. The temperature was increased to 110 °C for over-night. Using this procedure, the Vilsmeier reagent, Me2N+=CHCl CI", is generated in situ. The reaction was then cooled in an ice/water bath, quenched by adding ice and finally water (100 mL). The contents were cooled in an ice-water bath, then filtered and dried well to give 1.37 g (24%) desired product. 1H NMR (CDC13, 300 MHz): δ 10.51 (d, IH), 8.73 (d, IH, J= 0.6 Hz), 8.07 (d, IH, J= 5.7 Hz), 7.57 (dd, IH, J= 0.6 and 5.4 Hz); LCMS (m/z): 197 (MH*).. [0079] Compounds 11-14 were prepared by using above representative procedures of compounds 4-7. [0080] Analogously, the following compounds were made using above representative prodecures:
Figure imgf000031_0001
[0081] One of ordinary skill in the art would recognize that analogous chemistry can be done on heteroaryls analogous to compound 8, so long as the keto-functionality has an adjacent CH for ring annulation as outlined above. Examples of such starting materials would include, but not be limited to, the following optionally substituted heteroaryls:
Figure imgf000031_0002
Figure imgf000032_0001
Example 3 [0082] The compounds of the invention according to formula V can be synthesized according to the scheme III presented below using methods well known to those skilled in the art. One of ordinary skill in the art would recognize that analogous chemistry can be done on other fused heteroaryls analogous to compound la (below), so long as the nitrile- functionality can be installed on the "A" ring as outlined in Scheme III. As well, 2-halo (or other leaving group- containing) pyridines are commercially available and analogous heterocycles can be synthesized as outlined in Scheme II above using as starting materials, for example those listed above that are analogous to compound 8. For convenience, compounds 3a, 4a, 5a 6a and 7a are depicted as the corresponding 3H-tetrazole. One of ordinary skill in the art would recognize that, depending on reaction conditions, mixtures of the lH-tetrazole and the 3H-tetrazole are possible along with, less commonly, the 5H-tetrazole isomer. SCHEME III
Figure imgf000033_0001
Biological Assay
[0083] The following assay examples illustrate the HCV inhibitory properties of the compounds of the invention. Assay Example 1 HCVReplicon Assay [0084] Actively dividing 5-2Luc replicon cells were seeded at the density of 5000-7500 cells/well in the volume of 90 μl/well into 96 well plate(s). The cells were then incubated at 37°C and 5% CO2 for 24 hours. The 5-2 cells are replicon cells licensed from Ralf Bartenschlager (Germany) and have a self-replicating RNA molecule in the Huh7 cell; the RNA contains HCV non-structural proteins that make the self-replication possible.
[0085] Various concentrations of compounds (in the volume of 10 μl) were added into each well 24 hours after seeding the cells. The cells were incubated for another 24 hours before luciferase assay.
[0086] After incubating the 5-2Luc replicon cells with the compounds for 24 hours, media were aspirated from each well and Bright-Glo (Pharmacia) luciferase assay reagents were added to each well according to the manufacturer's manual. Briefly, the Bright-Glo reagent was diluted with equal volume of PBS and an aliquote (100 μl) was added to each well. After incubating the plate at room temperature for 5 minutes, luciferase counts were taken using a luminometer. Assay Example 2 Luciferase Counter Assay [0087] Actively dividing CMV-Luc cells (Luc cells in which DNA construct (CMV promoter followed by Luciferase gene) is permanently integrated into the chromosome of Huh7 cells) were seeded at the density of 5000-7500 cells/well in the volume of 90 μl/well into 96 well plate(s). The cells were then incubated at 37°C and 5% CO2 for 24 hours. [0088] Various concentrations of compounds (in the volume of 10 μl) were added into each well 24 hours after seeding the cells. The cells were incubated with the compounds for another 24 hours before luciferase assay.
[0089] After incubating the CMV-Luc cells with the compounds for 24 hours, media were aspirated from each well and Bright-Glo (Pharmacia) luciferase assay reagents were added to each well according to the manufacturer's manual. Luciferase counts were taken using a luminometer.
[0090] The activity of a number of compounds according to the invention measured by the luciferase assay is displayed in Table 1.
Assay Example 3 Immunoblotting Assay [0091] Actively dividing 9-13 replicon cells (Huh7 cells comprising an HCV replicon) were seeded at the density of lxl 05 cells/well in the volume of 2 ml/well into 6 well plate(s). The cells were then incubated at 37°C and 5% CO2 for 24 hours.
[0092] Various concentrations of compounds (in the volume of 10 μl) were added into each well 24 hours after seeding the cells. The cells were incubated with the compounds for another 48 hours.
[0093] Protein samples were prepared from the cultured cells and resolved on a SDS-PAGE gel. [0094] After electrophoresis, the protein samples on the SDS-PAGE gel were transferred to a nitrocellulose membrane.
[0095] The membrane was blocked with 5% non-fat milk in PBS for 1 hr at room temperature. [0096] Primary antibody incubation was performed for 1 hour at room temperature before the membrane was washed for 3 times with PBST (PBS plus 0.1% Tween 20), 15 minutes each. [0097] Horse Radish Peroxidase conjugated secondary antibody incubation was performed for 1 hour at room temperature before the membrane was washed for 3 times with PBST (PBS plus 0.1% Tween 20), 15 minutes each. [0098] The membrane was then soaked in substrate solution (Pierce) and exposed to a film.
Assay Example 4 TaqMan RT-PCR Assay
[0099] Actively dividing 9-13 replicon cells were seeded at the density of 3x104 cells/well in the volume of 1 ml/well into 24 well plate(s). The cells were then incubated at 37°C and 5% CO2 for
24 hours.
[0100] Various concentrations of compounds (in the volume of 10 μl) were added into each well 24 hours after seeding the cells. The cells were incubated with the compounds for another 24 hours.
[0101] After incubating the 9-13 replicon cells with the compounds for 24 hours, media were aspirated off and RNA samples were prepared from each well.
[0102] TaqMan® (Roche Molecular Systems) one step RT-PCR was performed using the RNA samples according to the manufacturer's manual. Briefly, properly diluted RNA sample, upstream primer, downstream primer, FAM-labeled probe oligo were mixed and water was added to make up the volume to 25 μl. Equal volume of 2X TaqMan Master Mix were added and the reaction was performed in an ABI Prism 7700 Sequence Detector (Applied Biosystems).
[0103] The compounds in Table 1 to 3 immediately below were prepared essentially using the methods described herein and illustrated in the schemes. All of the compounds in this application were named using Chemdraw Ultra version 6.0.2, which is available through Cambridgesoft.co, 100 Cambridge Park Drive, Cambridge, MA 02140, Namepro version 5.09, which is available from ACD labs, 90 Adelaide Street West, Toronto, Ontario, M5H, 3 V9, Canada, or were derived therefrom.
[0104] The compounds of Table 1 below exhibited greater than 20% inhibitory activity. Table 1
Figure imgf000036_0001
Figure imgf000037_0001
mpounds of Table 2 exhibited 5% to 20% inhibitory activity. TABLE 2
Figure imgf000037_0002
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
[00106] Compounds exhibiting less than 5% inhibitory activity are listed in Table 3 below. TABLE 3
Figure imgf000040_0003
Figure imgf000040_0002
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
[00107] The compounds in Table 4 below also exhibit HCN inhibitory activity. Table 4
Figure imgf000049_0001
+ means gre
[00108] The compounds in Table 5 below inhibit HCV activity from 4% to 110% at a concentration of 3 uM. Table 5
Figure imgf000049_0002
Figure imgf000050_0001

Claims

We claim:
1. A compound of formulae I or II,
Figure imgf000051_0001
II and pharmaceutically acceptable salts, hydrates, solvates andN-oxides thereof wherein, Ri is hydrogen, -OH, Ci-Ce-alkyl, heterocyclyl, Cι-C6-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, -CO-R7, Cι-C6- alkyl-NH-OR7, Cι-C6-alkyl-NH-OH, Cι-C6-alkyl-NR7-OH, Cι-C6-alkyl-NR7-OR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Cι-C6-alkyl- CO-NR7OH, -S02R7, -SOR7, -S02NH2, -S02NHR7, -SO2NR7R7, -SO2-heteroaryl, -S02-aryl, -SO3H, -SO3R7, -SO2Cl, -NHR7, Cι-C6-alkyl-NH(R7)-aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)- aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO- NH-(Cι-C6-alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Ci- C6-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl) or halogen; each of R2, R3, R4, R5 and R6 is independently selected from hydrogen, nitro, -OH, Cι-C6-alkyl, heterocyclyl, -0-halo(d-C6-alkyI), halo(Cι-C6-alkyl), d-Ce-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -OCF3, -CF3, azido, -O-(C3-C6-cycloalkyl), - S-(Cι-C6-alkyl), Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, - CO-R7, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH-OH, Cι-C6-alkyl-NR7-OH, Cι-C6-alkyl-NR7- OR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, C C6-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -SO2R7, -SOR7, -S02NH2, -SO2NHR7, -SO2NR7R7, -SO2- heteroaryl, -S02-aryl, -SO3H, -SO3R7, -SO2Cl, -NH2, -NHR7, -NR7R7, Cι.C6.alkyl-NH(R7)- aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)-aryl, -CO-heteroaryl, -N(R7)-CO-R7, -NH-S02-R7, - N(R7)-CO-NR7R7, -NH-CO-0-R7-aryl, -NH-CO-NH-S02-aryl, -NR7-CO-OR7, -NH-CO-OR7, -NH-CO-NH-(d-Ce-alkyl) or -NH-CO-(Cι-Ce-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-0-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι- C6-alkyl) or halogen; or any adjacent two of R3, R , R5 and R6 together with the carbon atoms to which they are attached form an aryl, heterocyclyl or heteroaryl; or
Ri and R2 are independently H or a group selected from
Figure imgf000052_0001
wherein X is =NH or =S, Y is =N-, R8 is hydrogen or hydroxy, R9 is hydroxy, Ci-Ce-alkoxy or aryl optionally substituted with hydroxy or Cι-C6-alkyl; or when adjacent, optionally Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl; each of Rio, Rn, Rι2 and Rι3 is independently selected from hydrogen, -CN, -N02, -OH, Ci-Ce- alkyl, aryl, Cι-C6-alkyl-aryl, -0-halo(Cι-C6-alkyl), halo(Cι-Ce-alkyl), -O-(C3-C6- cycloalkyl), -S-(Cι-C6-alkyl), heterocyclyl, Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-R7, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-NR7OH, -CO-NHOR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, d-C6-alkyl-CO-NR7OH, -CO- N(R7)OH, -CO-NHOH, -CO-H, -SO2-R7, -SO-R7, -SO-(Cι-Ce-alkyl), -SO2-(Cι-C6-alkyl), - SO2NH2, -S02NHR7, -S02NR7R7, -CO-heteroaryl, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH- OH Cι-C6-alkyl-NR7-OH Cι-C6-alkyl-NR7-OR7) -NH2, -NHR7, -NR7R7, -N(R7)-CO-R7, - NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7, or when the annular B ring atom to which it is bound is N, R3 is optionally absent, wherein each of the alkyl, alkoxy, aryl, heteroaryl and heterocyclyl are optionally substituted with one or more halogen, Cι-C6-alkyl, Cι-C6- alkoxy, -OCF3, -CF3, -CN, -NH2, -N02, -OH, mono- or di-Cι-C6-alkylamino, or oxo;
R7 is hydrogen, a pro-drug group, Co-C6-alkyl, -(Cι-Ce-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6- alkyl), -(Cι-C6-alkyl)-CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Ci-Ce-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, -CO-O-(Cι-C6-alkyl), cyano, -O-halo(Cι-C6- alkyl), halo(Cι-C6-alkyl) or halogen; Ai, A2, A3, and A4 are independently carbon, oxygen, nitrogen, sulfur, -S(O)-, -S(O2)-, -C(O)-, or -N(R7)-, wherein when Ai and A2 are carbon, then the group formed by Ai, A2 together with Rio and Ru is not phenyl; A5 is carbon or nitrogen; Bi, B2, B3, and B4 are independently a covalent bond, C, O, -S(O)o-2- or N, wherein when one of Bi, B2, B3, and B is a covalent bond the R group attached to it is absent and the B ring is a five membered ring, and when B is O, -S(O)o-2- or N then the R group attached is absent; and the dashed lines of rings A, B and C represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states; provided at least one of Bi, B2, B3 and B of Formula L is a heteroatom.
2. A compound according to claim 1 of the formula
Figure imgf000053_0001
and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein Ri and R2 are independently hydrogen, -OH, Ci-C6-alkyl, Cι-C6-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, halogen, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, -CO-R7, d-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH-OH, d-Ce-alkyl-NR7-OH, Cι-C6-alkyl-NR7-OR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2iιeteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2Cl, -SO2NHR7, -SO2N(R7), -NHR7, Cι-C6-alkyl-NH(R7)-aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)-aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO-NH-(Cι-Ce-alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Ci-Cβ-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι-C6- alkyl) or halogen; or one of Ri or R2 is H and the other is
Figure imgf000054_0001
wherein X is =NH or =S, Y is =N-, R8 is hydrogen or hydroxy, R9 is hydroxy, Ci-Ce-alkoxy or aryl optionally substituted with hydroxy or Ci-Ce-alkyl; or
Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl;
R3, R4, R5 and Re are independently hydrogen, nitro, Cι-C6-alkyl, aryl, -O-halo(d-C6-alkyl), halo(Ci-C6-alkyl), -O-(C3-C6-cycloalkyl), -S-(Cι-C6-a]kyl), -SO-(Cι-C6-alkyl), -SO2-(d-C6- alkyl), heterocyclyl, Ci-Cβ-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO- H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-R7, -CO- NHOH, -CO-NHOR7, -CO-NR7OH, -CO-NR7OR7, -SO2-(Cι-C6-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, -NH2, -NHR7, -NR7R7, -OH, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7, -N(R7)-CO-NR7R7, or, when the annular B ring atom to which it is bound is N, R3-R6 is optionally absent; or
R4 and R5 together with the carbon atoms to which they are attached form a heteroaryl;
R7 is hydrogen, a pro-drug group, Co-C6-alkyl, -(Cι-Ce-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6- alkyl), -(Cι-Ce-alkyl)-CN, d-Ce-alkene, heterocyclyl, aryl, heteroaryl or-(Ci-C6-alkyl)- aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Ci-Ce-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, -CO-O-(Cι-Ce-alkyl), cyano, -O-halo(Cι- Ce-alkyl), halo(Cι-C6-alkyl) or halogen;
Rio, Rπ, and R12 are independently hydrogen, -CN, -NO2, -OH, Ci-Ce-alkyl, aryl, Cι-C6-alkyl- aryl, -O-halo(d-C6-alkyl), halo(Cι-Ce-alkyl), -O-(C3-C6-cycloalkyl), -S-(Cι-C6-alkyl), -SO- (Ci-Ce-alkyl), -SO2-(Cι-C6-alkyl), heterocyclyl, Ci-Ce-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-R7, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-NHOR7, Ci-C6-alkyl-CO-NHOR7, d-C6-alkyl-CO-NR7OR7, Ci-Ce-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -CO-N(R7)OH, -CO-NHOH, -CO-H, -SO2-R7, -SO-R7, -SO2-(Cι-C6-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, Ci-Ce-alkyl-NH-OR?, Cι-C6-a]kyl-NH-OH Cι-C6-alkyl-NR7-OH Cι-C6-a.kyl-NR7-OR7j -NH2, -NHR7, -NR7R7, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7, wherein each of the alkyl, alkoxy, aryl, heteroaryl and heterocyclyl are optionally substituted with one or more halogen, Ci-Ce-alkyl, Cι-C6-alkoxy, -OCF3, -CF3, -CN, -NH2, -NO2, -OH, mono- or di-Cι-C6-alkylamino, or oxo;
Ai, A2, and A3 are independently carbon, oxygen, nitrogen, sulfur, -S(O)-, -S(O2)-, -C(O)-, or -N(R7)-;
A4 is carbon or nitrogen;
Bi, B2, B3, and B are independently a covalent bond, C, O, -S(O)0-2- or N, wherein when one of Bi, B2, B3, and B is a covalent bond the R group attached to it is absent and the B ring is a five membered ring; and the dashed lines of rings B and A represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states; provided at least one of Bi, B2, B3 and B is a heteroatom.
3. A compound according to claim 2 of the formula
Figure imgf000055_0001
and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein Ri and R2 are independently hydrogen, Cι-C6-alkyl, heterocyclic, Cι-C6-alkyl-OH, aryl, heteroaryl, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO- NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-N(R7)-OR7, -CO-R7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -SO3H, -NHR7, Cι-C6-alkyl-NH(R7)-aryl, -NH(R7)-aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO- OR7, -NH-CO-NH-(Cι-Ce-alkyl) or -NH-CO-(Cι-Ce-alkyl), wherein each of the heteroaryl and aryl groups are optionally substituted with Ci-Cβ-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-C6-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(C Ce-alkyl) or halogen; or Ri or R2 is a group selected from
Figure imgf000056_0001
wherein R8 is hydrogen or hydroxy; X is =NH or =S, Y is =N-, and R9 is hydroxy, Ci-Ce-alkoxy or aryl optionally substituted with hydroxy or Ci-Ce-alkyl; or
Ri and R2 together with the carbon atoms to which they are attached form a mono or bicychc aryl or heteroaryl;
R3, R4, R5 and Re are independently hydrogen, nitro, Ci-Ce-alkyl, aryl, -O-halo(Ci-Ce-alkyl), halo(Cι-Ce-alkyl), -O-(C3-C6-cycloalkyl), -S-(Cι-C6-alkyl), heterocyclyl, Cι-C6- alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-OH, -CO- OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-R7, -SO3H, -SO2-(Cι- C6-alkyl), -SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, -NH2, -NHR7, -NR7R7, -OH, -N(R7)-CO-R7, -NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7;
R and R5 together with the carbon atoms to which they are attached form a heteroaryl;
R7 is hydrogen, C0-C6-alkyl, -(Cι-C6-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6-alkyl), -(Cι-C6-alkyl)- CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Cι-C6- alkyl, nitro, hydroxy, Ci-C6-alkoxy, -CO-O-(Ci-Ce-alkyl), cyano, -O-halo(Cι-C6- alkyl), halo(Cι-C6-alkyl) or halogen;
Ai, A2 and A3 are independently carbon, oxygen, or nitrogen; and ring B contains 1 or 2 nitrogen atoms.
4. The compound according to claim 3, wherein Ri is -CO-NH-OR7, R5 is halogen, and 2 to R4 and e are hydrogen.
5. The compound according to claim 4, wherein R7 is hydrogen and R5 is chloro.
6. The compound according to claim 3, wherein Ri is aryl optionally substituted with one or more halo(Cι-C6-alkyl), cyano, -O-halo(Cι-Ce-alkyl), or halogen, R5 is halogen, R to Ri and Re are hydrogen.
7. The compound according to claim 6 wherem Ri is phenyl optionally substituted with one or more -CF3 or F and R5 is chloro.
8. The compound according to claim 5, wherein Ri is -CO-NH-OR7, Re is halogen and R2 to R5 are hydrogen.
9. The compound according to claim 19, wherein R7 is isobutyl, propyl, isopropyl or ethyl and Re is chloro.
10. The compound according to claim 5, wherein Ri is -NH-CO-OR , R5 is halogen, R2 to R and R are hydrogen.
11. The compound according to claim 10 wherein R7 is tert-butyl and R5 is chloro.
12. The compound according to claim 3, wherein Ri is -CO-OR , Re is halo(Cι-Ce-alkyl) and R2 to R5 are hydrogen.
13. The compound according to claim 12, wherein R7 is hydrogen and R is -CF3.
14. The compound according to claim 3, wherein Ri is -NH-CO-NH-Sθ2-aryl, -NH-CO-NH-(Cι-Ce-alkyl), Ci-Ce-alkyl-OH, -CO-H, -CO-NH2, -CO-NH-R7, -CO-NH- OR7, -Sθ2-aryl, -Sθ2-heteroaryl, heteroaryl, -CO-OR7 or a group represented by
Figure imgf000057_0001
wherein R8 is hydrogen or hydroxy, X is =N or =S, Y is -N-, R9 is hydroxy or Ci-Ce- alkoxy; R4 and R5 are independently selected from hydrogen, -S-(Cι-Ce-alkyl), Ci-Cβ- alkyl, -O-(C3-C6-cycloalkyl), Ci-Ce-alkoxy, halogen, -SO2-(Cι-C6-alkyl), heterocyclyl, -O-halo(Cι-C6-alkyl), halo(Cι-Ce-alkyl), -NH2 or together with the carbon atom to which they are attached form a heteroaryl group; R2, R3 and Re are hydrogen; and wherein the each of the aryl, heteroaryl and heterocyclyl groups are optionally substituted with -CO- O-(Cι-Ce-alkyl), Ci-Ce-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, cyano, -O-halo(Cι-Ce-alkyl), halo(Cι-Ce-alkyl) or halogen.
15. The compound according to claim 3, wherein Ri and R2 are independently selected from is -NH(R7)-aryl, -CO-OH, -CO-OR7, Cι-C6-alkyl, aryl or together with the carbon atoms to which they are attached form a bicyclic heteroaryl; R3 to Re are hydrogen; and wherein each of the aryl and heteroaryl groups are optionally substituted with -CO-O-(Cι-C6- alkyl), Ci-Ce-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, cyano, -O-halo(Cι-C6-alkyl), halo(Cι- C6-alkyl) or halogen.
16. The compound according to claim 3, wherein Ri is -CO-OH, -CO-OR7, or -CO-NH-OR7; Re is Ci-Ce-alkyl, halogen, or -O-(Cι-Ce-alkyl); and R2 to R5 are hydrogen.
17. The compound according to claim 3, wherein Ri is -CO-OR7; R3 is azido or Ci-Ce-alkyl; and R2, R4 to Re are hydrogen.
18. The compound according to claim 2 wherein one of Bi, B2, B3, and B is a covalent bond, and another is S and the remaining two are C, wherein the dashed lines of ring B are independently single or double bonds such that the annular S has two bonds and each annular C has four bonds; Ai, A2, A3 and A are independently carbon or nitrogen; R2 to Re and Rio to R12 are hydrogen; Ri is
Figure imgf000058_0001
wherein R9 is hydroxy or C C3-alkoxy, -CO-H, -CO-OH, -CO-NH-OH, or -CO-NH- OR7, wherein R7 is a heterocyclyl, wherein the heterocyclyl groups is optionally substituted with Cι-C6-alkyl, hydroxy, Ci-Ce-alkoxy, cyano, or halogen.
19. The compound according to claim 18, wherein Ai to A4 are nitrogen.
20. The compound according to claim 19, wherein Bi is S, B2 and B3 are C and B4 is a covalent bond.
21. The compound according to claim 20, wherein the R is methoxy.
22. The compound according to claim 20, wherein the heterocyclyl is pyranyl.
23. The compound according to claim 22, wherem the heterocyclyl is pyran-2-yl.
24. The compound according to claim 20 that is selected from the group consisting of tefrazolo[l,5-a]tlιieno[2,3-e]pyridine-4-carboxylic acid; N-(tefrahydro-2H-pyran-2-yloxy)tefrazolo[l,5-a]t eno[2,3-e]pyridine-4-carboxamide; N-hydroxytefrazolo[l,5-a]tMeno[2,3-e]pyridine-4-carboxamide; tefrazolo[l,5-α]tMeno[2,3-e]pyridine-4-carbaldehyde; tefrazolo[l,5-α]thieno[2,3-e]pyridine-4-carbaldehyde O-methyloxime; and and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof.
25. A compound according to claim 1 of the formula
Figure imgf000059_0001
and pharmaceutically acceptable salts, hydrates, solvates and N-oxides thereof wherein Ri is hydrogen, -OH, Ci-Ce-alkyl, heterocyclyl, Cι-C6-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, -CO-R7, C C6- alkyl-NH-OR7, Ci-Ce-alkyl-NH-OH, Cι-Ce-alkyl-NR7-OH, Cι-C6-alkyl-NR7-OR7, Cι-C6-alkyl-CO-NHOR7, Cι-C6-a]kyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Ci-Ce-alkyl- CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -SO2NR7R7, -SO2-heteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2CI, -NHR7, Cι-Ce-alkyl-NH(R7)-aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)- aryl, -CO-heteroaryl, -NH-CO-O-R7-aryl, -NH-CO-NH-SO2-aryl, -NH-CO-OR7, -NH-CO- NH-(Cι-C6-alkyι) or -NH-CO-(Cι-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Ci- C6-alkoxy, -CO-0-(Cι-C6-akyl), cyano, -O-halo(Cι-Ce-alkyl), halo(Cι-C6-alkyl) or halogen; each of R2, R3, R , R5 and R6 is independently selected from hydrogen, nitro, -OH, Ci-Ce-alkyl, heterocyclyl, -O-halo(Cι-C6-alkyl), halo(Cι-C6-alkyl), Cι-C6-alkyl-OR7, Cι-C6-alkyl-OH, aryl, heteroaryl, heterocyclic, halogen, cyano, -OCF3, -CF3, azido, -O-(C3-C6-cycloalkyl), - S-(Cι-C6-alkyl), Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, -CO-H, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-NH-OH, -CO-NH-OR7, -CO-NR7-OH, -CO-N(R7)-OR7, - CO-R7, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH-OH, Cι-C6-alkyl-NR7-OH, Cι-C6-alkyl-NR7- OR7, Cι-C6-alkyl-CO-NHOR7, C C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Cι-C6-alkyl-CO-NR7OH, -SO2R7, -SOR7, -SO2NH2, -SO2NHR7, -S02NR7R7, -S02- heteroaryl, -SO2-aryl, -SO3H, -SO3R7, -SO2Cl, -NH2, -NHR7, -NR7R7, d.C6-alkyl-NH(R7)- aryl, Cι-C6-alkyl-NR7-OR7, -NH(R7)-aryl, -CO-heteroaryl, -N(R7)-CO-R7, -NH-S02-R7, - N(R7)-CO-NR7R7, -NH-CO-O-R7-aryl, -NH-CO-NH-S02-aryl, -NR7-CO-OR7, -NH-CO-OR7, -NH-CO-NH-(Cι-Ce-alkyl) or -NH-CO-(Cι-C6-alkyl), wherein each of the alkyl, heterocyclyl, heteroaryl and aryl groups are optionally substituted with Cι-C6-alkyl, nitro, hydroxy, Cι-C6-alkoxy, -CO-O-(Cι-Ce-akyl), cyano, -O-halo(Cι-C6-alkyl), halo(Cι- C6-alkyl) or halogen; or any adjacent two of R3, R , R5 and R6 together with the carbon atoms to which they are attached form an aryl, heterocyclyl or heteroaryl; or
Ri and R2 are independently H or a group selected from
Figure imgf000060_0001
wherein X is =NH or =S, Y is =N-, R8 is hydrogen or hydroxy, R9 is hydroxy, Ci-C6-alkoxy or aryl optionally substituted with hydroxy or Ci-Ce-alkyl; or when adjacent, optionally Ri and R2 together with the carbon atoms to which they are attached form a mono or bicyclic aryl or heteroaryl; each of Rio, Rπ, Rι2 and R13 is independently selected from hydrogen, -CN, -N02, -OH, C1-C6- alkyl, aryl, Cι-C6-alkyl-aryl, -O-halo(Cι-C6-alkyl), halo(Cι-Ce-alkyl), -O-(C3-C6- cycloalkyl), -S-(Cι-C6-alkyl), heterocyclyl, Cι-C6-alkoxy, -(Cι-C6-alkoxy)-aryl, azido, halogen, -OCF3, -CF3, -CO-H, -CO-R7, -CO-OH, -CO-OR7, -CO-NH2, -CO-NHR7, -CO-NR7R7, -CO-N(R7)OR7, -CO-NR7OH, -CO-NHOR7, Cι-C6-alkyl-CO-NHOR7, Ci-C6-alkyl-CO-NR7OR7, Cι-C6-alkyl-CO-NHOH, Ci-C6-aucyl-CO-NR7OH, -CO- N(R7)OH, -CO-NHOH, -CO-H, -SO2-R7, -SO-R7, -SO-(Cι-C6-alkyl), -SO2-(Cι-C6-alkyl), - SO2NH2, -SO2NHR7, -SO2NR7R7, -CO-heteroaryl, Cι-C6-alkyl-NH-OR7, Cι-C6-alkyl-NH- OHj Cι-Ce-a]kyl-NR7-OH) Cι-C6-alkyl-NR7-OR7>-NH2, -NHR7, -NR7R7, -N(R7)-CO-R7, - NHSO2R7, -N(R7)-CO-OR7 or -N(R7)-CO-NR7R7, or when the annular B ring atom to which it is bound is N, R3 is optionally absent, wherein each of the alkyl, alkoxy, aryl, heteroaryl and heterocyclyl are optionally substituted with one or more halogen, Cι-C6-alkyl, Cι-C6- alkoxy, -OCF3, -CF3, -CN, -NH2, -NO2, -OH, mono- or di-Cι-C6-alkylamino, or oxo;
R7 is hydrogen, a pro-drug group, Co-C6-alkyl, -(Cι-Ce-alkyl)-OH, -(Cι-C6-alkyl)-O-(Cι-C6- alkyl), -(Cι-C6-alkyl)-CN, C2-C6-alkene, heterocyclyl, aryl, heteroaryl or -(Cι-C6-alkyl)-aryl, wherein each of the aryl, heterocyclyl and heteroaryl groups are optionally substituted with Ci-Cβ-alkyl, nitro, hydroxy, Ci-Ce-alkoxy, -CO-O-(Ci-C6-alkyl), cyano, -O-halo(Cι-C6- alkyl), halo(Cι-C6-alkyl) or halogen;
A], A2, A3, and A4 are independently carbon, oxygen, nifrogen, sulfur, -S(O)-, -S(O2)-, -C(O)-, or -N(R7)-, wherein when Ai and A2 are carbon, then the group formed by Ai, A2 together with Rio and Rn is not phenyl;
A5 is carbon or nitrogen;
Bi, B2, B3, and B4 are independently a covalent bond, C, O, -S(O)0-2- or N, wherein when one of Bi, B2, B3, and B4 is a covalent bond the R group attached to it is absent and the B ring is a five membered ring, and when B is O, S or N then the R group attached is absent; and the dashed lines of rings A, B and C represent single or double bonds such that each annular N has three bonds, each annular O has two bonds, each annular S has two bonds and each annular C has four bonds; annular sulfur atoms may exist as sulfide, sulfoxide and sulfone oxidation states.
26. A composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.
27. A method of inhibiting HCV proliferation comprising contacting an HCV infected cell with a compound according to claim 1.
28. A method of treating a mammal infected with an HCV infection, the method comprising administering to the mammal a therapeutically effective amount of a composition according to claim 26.
29. The method according to claim 28 wherein the mammal is a human.
PCT/US2005/019499 2004-06-03 2005-06-03 Heterotricyclic compounds for use as hcv inhibitors WO2005121138A2 (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US57653504P 2004-06-03 2004-06-03
US57652104P 2004-06-03 2004-06-03
US60/576,535 2004-06-03
US60/576,521 2004-06-03
US64521205P 2005-01-18 2005-01-18
US60/645,212 2005-01-18

Publications (2)

Publication Number Publication Date
WO2005121138A2 true WO2005121138A2 (en) 2005-12-22
WO2005121138A3 WO2005121138A3 (en) 2009-03-05

Family

ID=35503711

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2005/019499 WO2005121138A2 (en) 2004-06-03 2005-06-03 Heterotricyclic compounds for use as hcv inhibitors

Country Status (2)

Country Link
US (1) US7569580B2 (en)
WO (1) WO2005121138A2 (en)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11492365B2 (en) 2020-02-07 2022-11-08 Gasherbrum Bio, Inc. Heterocyclic GLP-1 agonists
US11661422B2 (en) 2020-08-27 2023-05-30 Incyte Corporation Tricyclic urea compounds as JAK2 V617F inhibitors
US11691971B2 (en) 2020-06-19 2023-07-04 Incyte Corporation Naphthyridinone compounds as JAK2 V617F inhibitors
US11753413B2 (en) 2020-06-19 2023-09-12 Incyte Corporation Substituted pyrrolo[2,1-f][1,2,4]triazine compounds as JAK2 V617F inhibitors
US11767323B2 (en) 2020-07-02 2023-09-26 Incyte Corporation Tricyclic pyridone compounds as JAK2 V617F inhibitors
US11780840B2 (en) 2020-07-02 2023-10-10 Incyte Corporation Tricyclic urea compounds as JAK2 V617F inhibitors
US11919908B2 (en) 2020-12-21 2024-03-05 Incyte Corporation Substituted pyrrolo[2,3-d]pyrimidine compounds as JAK2 V617F inhibitors
US11958861B2 (en) 2021-02-25 2024-04-16 Incyte Corporation Spirocyclic lactams as JAK2 V617F inhibitors

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2287466T3 (en) * 2002-03-22 2007-12-16 Janssen Pharmaceutica N.V. DERIVATIVES OF 2-QUINOLINONA AND QUINAZOLINA FROM BENCILIMIDAZOLIL SUBSTITUTED AS INHIBITORS OF FARNESIL TRANSFERASA.
US10513515B2 (en) 2017-08-25 2019-12-24 Biotheryx, Inc. Ether compounds and uses thereof
CA3106239A1 (en) 2018-07-27 2020-01-30 Biotheryx, Inc. Bifunctional compounds as cdk modulators
US11897930B2 (en) 2020-04-28 2024-02-13 Anwita Biosciences, Inc. Interleukin-2 polypeptides and fusion proteins thereof, and their pharmaceutical compositions and therapeutic applications

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2206012A1 (en) * 1971-02-11 1972-08-24 Aspro-Nicholas Ltd., Slough, Buckinghamshire (Grossbritannien) Pharmaceutical preparation
GB1374369A (en) * 1971-08-16 1974-11-20 Lilly Co Eli Triazoloquinoline derivatives and their use as agents for the control of plant-pathogenic organisms
US4000151A (en) * 1974-09-11 1976-12-28 The Upjohn Company Triazolyl benzophenone compounds
EP0120484A1 (en) * 1983-03-25 1984-10-03 Merrell Dow Pharmaceuticals Inc. (1H-tetrazol-5-yl)tetrazolo (1,5-a)-quinolines and related compounds
WO1995026348A1 (en) * 1994-03-25 1995-10-05 Merrell Dow Pharmaceuticals Inc. PROCESS FOR PREPARING (1H-TETRAZOL-5-yl)TETRAZOLO [1,5-a] QUINOLINES AND NAPHTHYRIDINES
WO1997048704A1 (en) * 1996-06-21 1997-12-24 Minnesota Mining And Manufacturing Company Process for preparing imidazoquinolinamines
US20040082592A1 (en) * 2000-10-02 2004-04-29 Mabire Dominique Jean-Pierre Metabotropic glutamate receptor antagonists
WO2005030774A1 (en) * 2003-09-26 2005-04-07 Rigel Pharmaceuticals, Inc. Hcv inhibitors and methods of using them

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4581455A (en) * 1984-10-10 1986-04-08 Merrell Dow Pharmaceuticals Inc. Tetrazolothienopyridines
ID30204A (en) 1999-12-27 2001-11-15 Japan Tobacco Inc COMPOUNDS OF DIFFUSED RING AND ITS USE AS A MEDICINE
EP1322636A1 (en) 2000-09-25 2003-07-02 Janssen Pharmaceutica N.V. Farnesyl transferase inhibiting 6- (substituted phenyl)methyl]-quinoline and quinazoline derivatives
JP4558314B2 (en) 2001-07-20 2010-10-06 ベーリンガー インゲルハイム （カナダ） リミテッド Viral polymerase inhibitor
MY151199A (en) 2001-11-02 2014-04-30 Rigel Pharmaceuticals Inc Substituted diphenyl heterocycles useful for treating hcv infection
CA2370396A1 (en) 2002-02-01 2003-08-01 Boehringer Ingelheim (Canada) Ltd. Hepatitis c inhibitor tri-peptides

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2206012A1 (en) * 1971-02-11 1972-08-24 Aspro-Nicholas Ltd., Slough, Buckinghamshire (Grossbritannien) Pharmaceutical preparation
GB1374369A (en) * 1971-08-16 1974-11-20 Lilly Co Eli Triazoloquinoline derivatives and their use as agents for the control of plant-pathogenic organisms
US4000151A (en) * 1974-09-11 1976-12-28 The Upjohn Company Triazolyl benzophenone compounds
EP0120484A1 (en) * 1983-03-25 1984-10-03 Merrell Dow Pharmaceuticals Inc. (1H-tetrazol-5-yl)tetrazolo (1,5-a)-quinolines and related compounds
WO1995026348A1 (en) * 1994-03-25 1995-10-05 Merrell Dow Pharmaceuticals Inc. PROCESS FOR PREPARING (1H-TETRAZOL-5-yl)TETRAZOLO [1,5-a] QUINOLINES AND NAPHTHYRIDINES
WO1997048704A1 (en) * 1996-06-21 1997-12-24 Minnesota Mining And Manufacturing Company Process for preparing imidazoquinolinamines
US20040082592A1 (en) * 2000-10-02 2004-04-29 Mabire Dominique Jean-Pierre Metabotropic glutamate receptor antagonists
WO2005030774A1 (en) * 2003-09-26 2005-04-07 Rigel Pharmaceuticals, Inc. Hcv inhibitors and methods of using them

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
FILER C N ET AL: "ISOQUINOLINES. 8. ETHYLENE OXIDE MEDIATED CONVERSION OF ISOQUINOLINES TO ISOQUINOLONES AND OXAZOLIDINES. ITS EXTENSION TO RELATED NITROGEN HETEROCYCLES" 1979, JOURNAL OF ORGANIC CHEMISTRY, AMERICAN CHEMICAL SOCIETY. EASTON, US, PAGE(S) 285-287 , XP002313578 ISSN: 0022-3263 page 286; compound 7 *
NISPEN VAN S P J M ET AL: "USE OF DILITHIO-TOSYLMETHYL ISOCYANIDE IN THE SYNTHESIS OF OXAZOLES AND IMIDAZOLES" TETRAHEDRON LETTERS, ELSEVIER, AMSTERDAM, NL, vol. 21, 1980, pages 3723-3726, XP002313580 ISSN: 0040-4039 *
ROBERTS E M ET AL: "A SYNTHESIS OF 5,6-BENZOPYRROCOLINE" JOURNAL OF ORGANIC CHEMISTRY, AMERICAN CHEMICAL SOCIETY. EASTON, US, vol. 20, 1955, pages 1443-1447, XP002313582 ISSN: 0022-3263 *
SMITH R M ET AL: "STRUCTURE-BASED DESIGN OF HEPATITS C VIRUS INHIBITORS" JOURNAL OF VIRAL HEPATITIS, BLACKWELL, OXFORD, GB, vol. 10, 2003, pages 405-412, XP002313583 ISSN: 1352-0504 *
TOMINAGA Y ET AL: "POLARIZED ETHYLENE. V. SYNTHESIS OF 1-SUBSTITUTED INDOLIZINE, PYRAZOLO[1,5-A]PYRIDINE, AND THEIR RELATED COMPOUNDS USING METHOXYETHYLENE DERIVATIVES" JOURNAL OF HETEROCYCLIC CHEMISTRY, HETEROCORPORATION. PROVO, US, vol. 27, no. 263, 1990, pages 263-268, XP002313581 ISSN: 0022-152X *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11492365B2 (en) 2020-02-07 2022-11-08 Gasherbrum Bio, Inc. Heterocyclic GLP-1 agonists
US11926643B2 (en) 2020-02-07 2024-03-12 Gasherbrum Bio, Inc. Heterocyclic GLP-1 agonists
US11691971B2 (en) 2020-06-19 2023-07-04 Incyte Corporation Naphthyridinone compounds as JAK2 V617F inhibitors
US11753413B2 (en) 2020-06-19 2023-09-12 Incyte Corporation Substituted pyrrolo[2,1-f][1,2,4]triazine compounds as JAK2 V617F inhibitors
US11767323B2 (en) 2020-07-02 2023-09-26 Incyte Corporation Tricyclic pyridone compounds as JAK2 V617F inhibitors
US11780840B2 (en) 2020-07-02 2023-10-10 Incyte Corporation Tricyclic urea compounds as JAK2 V617F inhibitors
US11661422B2 (en) 2020-08-27 2023-05-30 Incyte Corporation Tricyclic urea compounds as JAK2 V617F inhibitors
US11919908B2 (en) 2020-12-21 2024-03-05 Incyte Corporation Substituted pyrrolo[2,3-d]pyrimidine compounds as JAK2 V617F inhibitors
US11958861B2 (en) 2021-02-25 2024-04-16 Incyte Corporation Spirocyclic lactams as JAK2 V617F inhibitors

Also Published As

Publication number Publication date
US7569580B2 (en) 2009-08-04
WO2005121138A3 (en) 2009-03-05
US20050282850A1 (en) 2005-12-22

Similar Documents

Publication Publication Date Title
WO2005121138A2 (en) Heterotricyclic compounds for use as hcv inhibitors
US20050215614A1 (en) Substituted indoles and their use as hcv inhibitors
US10647711B2 (en) Azepin-2-one derivatives as RSV inhibitors
ES2808987T3 (en) Aryl and Heteroaryl Fused Lactams
US7868024B2 (en) Derivatives of N-(heteroaryl)-1-heteroaryl-1H-indole-2-carboxamides, preparation thereof and therapeutic use thereof
ES2326750T3 (en) 5-CARBOXIAMIDO PIRAZOLES AND (1,2,4) TRIAZOLS AS ANTIVIRAL AGENTS.
JPS62201882A (en) Isoflavon derivative
BG107285A (en) Beta-carboline derivatives useful as inhibitors of phosphodiesterase
US20130303747A1 (en) Pyrimidine nucleoside derivatives, synthesis methods and uses thereof for preparing anti-tumor and anti-virus medicaments
EA034235B1 (en) Novel compositions, uses and methods for making them
JP2011503196A (en) 3-Aminosulfonyl-substituted indole derivatives and methods for their use
EP0319429B1 (en) 9-Acylamino-tetrahydroacridine derivatives and memory enhancing agent containing said derivative as active ingredient
WO2018177296A1 (en) Lactam compound, and preparation method therefor and use thereof
CN111801100A (en) 7-substituted sulfoxy-purinone compounds and derivatives for the treatment and prevention of liver cancer
US20070149520A1 (en) HCV Inhibitors And Methods Of Using Them
WO2022100570A1 (en) Sarm1 enzyme activity inhibitor and use thereof in neurodegenerative diseases
WO2005058315A1 (en) Novel heterocyclic compounds as ikk2 inhibitors with anti-hbv activity
KR20040077735A (en) Tricyclic 2-Pyrimidone Compounds Useful as HIV Reverse Transcriptase Inhibitors
WO2018028491A1 (en) Indoleamine2,3-dioxygenase inhibitors and uses thereof in pharmacy
JPH04305578A (en) Benzodiazepine, method of manufacturing same and use thereof as pharmaceutical
JP2007506788A5 (en)
CN117343064B (en) Preparation and application of pyrimidine derivative with antiviral effect
CN113831346B (en) Multi-target anti-tumor small molecule and derivative, preparation method, pharmaceutical composition and application thereof
EP1814861A1 (en) Substituted 5-oxo pyrazoles and [1,2,4]triazoles as antiviral agents
CN108201539A (en) The application of biphenyl nucleosides phosphoramidate compounds

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: DE

WWW Wipo information: withdrawn in national office

Country of ref document: DE

122 Ep: pct application non-entry in european phase